Technical Field
[0001] The present invention relates to an EGFR inhibitor containing a pyrimidine compound
or a salt thereof.
Background Art
[0002] EGFR (which is also referred to as "ErbB 1 ") is receptor tyrosine kinase belonging
to the ErbB family and has been reported to contribute to cell growth or apoptosis
inhibition by binding to epidermal growth factor (which is also referred to as "EGF"),
mainly in epithelial tissues, among normal tissues (Non Patent Literature 1).
[0003] EGFR is considered to be a proto-oncogene. It has been reported that EGFR gene amplification,
overexpression, mutation and the like occur in various types of cancers. From non-clinical
and clinical research data, it is considered that activation of EGFR and downstream
signals play an important role in the survival and/or proliferation, etc. of cancer
cells associated with the genetic abnormality, overexpression and the like of EGFR.
For example, a mutation in exon 19 region of EGFR, which deletes amino acids at positions
746 to 750 (which is also referred to as "exon 19 deletion mutation"), and a mutation
in exon 21 region, which substitutes an amino acid leucine at position 858 with arginine
(which is also referred to as "L858R mutation") are considered to contribute to the
survival and/or proliferation of cancer cells by autophosphorylating EGFR in an EGF-independent
manner and thereby inducing kinase activity (Non Patent Literature 2). It has been
reported that these mutations are present in approximately 30 to 50% of non-small
cell lung cancer cases in East Asia and approximately 10% of non-small cell lung cancer
cases in Europe and the United States (Non Patent Literature 3).
[0004] Accordingly, an inhibitor capable of regulating the kinase activity of EGFR is assumed
to inhibit EGFR and downstream signals in cancer cells having EGFR gene amplification,
overexpression and/or mutation, so as to exhibit antitumor effects on the cancer cells.
Therefore, such an inhibitor is considered to be useful for the treatment, life-prolonging,
or QOL improvement of cancer patients.
[0005] Hence, a plurality of EGFR inhibitors have heretofore been researched and developed
as anticancer agents and are used in the treatment of EGFR mutation-positive tumor.
For example, drugs such as afatinib, gefitinib, and erlotinib have been approved as
therapeutic agents for EGFR mutation-positive lung cancer having exon 19 deletion
mutation or L858R mutation. Also, osimertinib has been approved as a therapeutic agent
for EGFR mutation-positive lung cancer having exon 19 deletion mutation or L858R mutation
as well as a mutation in exon 20 region, which substitutes an amino acid threonine
at position 790 with methionine (which is also referred to as "T790M mutation").
[0006] A mutation in exon 20 region, which insets one or more amino acids (which is also
referred to as "exon 20 insertion mutation") is also considered as an activating mutation
in lung cancer and the like (Non Patent Literature 4). It has been reported that cancer
having such a mutation tends to be low sensitive to a plurality of existing EGFR inhibitors.
For example, as for the clinical effect of afatinib on EGFR mutation-positive lung
cancer, it has been reported that its effect on exon 20 insertion mutation tends to
be much lower than that on exon 19 deletion mutation or L858R mutation (Non Patent
Literature 5). Regarding lung cancer having exon 20 insertion mutation of EGFR, multiple
clinical trials have been carried out. However, under the current circumstances, a
therapeutic method therefor has not yet been established. Therefore, it has been desired
to develop an EGFR inhibitor having inhibitory activity against exon 20 insertion
mutation.
[0007] It has been reported that brain metastasis occurs in approximately 25% to 40% of
lung cancer cases, in approximately 15% to 30% of breast cancer cases, and in certain
percentages of other multiple cancer cases (Non Patent Literatures 6 and 7).
[0008] From the viewpoint of the control of pathological conditions including brain metastasis
nidus, it has been desired to develop an EGFR inhibitor having inhibitory activity
against EGFR mutations and also having brain penetration properties.
Citation List
Non Patent Literature
[0009]
[Non Patent Literature 1] Nat. Rev. Cancer, vol. 6, pp. 803-812 (2006)
[Non Patent Literature 2] Nature Medicine, vol. 19, pp. 1389-1400 (2013)
[Non Patent Literature 3] Nat. Rev. Cancer, vol. 7, pp. 169-181 (2007)
[Non Patent Literature 4] Signal Transduct Target Ther. 4: 5, pp. 1-10 (2019)
[Non Patent Literature 5] Lancet Oncol. vol. 16, pp. 830-838 (2015)
[Non Patent Literature 6] Current Oncology, 25, p. S103-S114 (2018)
[Non Patent Literature 7] Breast Cancer Research, 18 (1), 8, p. 1-9 (2016)
Summary of Invention
Technical Problem
[0010] An object of the present invention is to provide a therapeutic agent for a disease
associated with EGFR, comprising a compound that has EGFR inhibitory activity and
has brain penetration properties as an active ingredient.
Solution to Problem
[0011] As a result of intensive studies, the present inventors have found that a compound
represented by the following formula (I) having pyrimidine as a basic skeleton, or
a salt thereof has EGFR inhibitory activity and brain penetration properties and is
useful as a therapeutic agent for diseases associated with EGFR (particularly, malignant
tumor) by inhibiting EGFR, reaching the completion of the present invention.
[0012] Specifically, one embodiment of the present invention includes the following:
- [1] A therapeutic agent for a disease associated with EGFR, comprising a pyrimidine
compound represented by the following formula (I), or a pharmaceutically acceptable
salt thereof as an active ingredient:
wherein R1 represents a C1-C4 alkyl group optionally having a C1-C4 alkoxy group as a substituent,
or a C3-C4 cycloalkyl group;
R2 represents a hydrogen atom, a halogen atom, a C1-C6 alkyl group optionally having
1 to 5 C1-C4 alkoxy groups or fluorine atoms each as a substituent(s), or a C1-C6
alkoxy group;
R3 represents a hydrogen atom, or a C1-C4 alkyl group optionally having 1 to 5 fluorine
atoms as a substituent(s);
R4 represents a hydrogen atom or a C1-C4 alkyl group; and
R5 represents a phenyl group optionally having 1 to 3 substituents selected from fluorine
atoms and chlorine atoms.
- [2] The therapeutic agent according to [1], wherein the pyrimidine compound is a compound
represented by the following formula (II):
wherein R1 represents a C1-C4 alkyl group optionally having a C1-C4 alkoxy group as a substituent,
or a C3-C4 cycloalkyl group;
R2 represents a hydrogen atom, a halogen atom, a C1-C6 alkyl group optionally having
1 to 5 C1-C4 alkoxy groups or fluorine atoms each as a substituent(s), or a C1-C6
alkoxy group;
R3 represents a hydrogen atom, or a C1-C4 alkyl group optionally having 1 to 5 fluorine
atoms as a substituent(s);
R4 represents a hydrogen atom or a C1-C4 alkyl group; and
R5 represents a phenyl group optionally having 1 to 3 substituents selected from fluorine
atoms and chlorine atoms.
- [3] The therapeutic agent according to [1] or [2], wherein the pyrimidine compound
is a compound represented by the formula (I) or (II) wherein
R2 is a C1-C6 alkyl group optionally having 1 to 5 C1-C4 alkoxy groups as a substituent(s).
- [4] The therapeutic agent according to any of [1] to [3], wherein the pyrimidine compound
is a compound represented by the formula (I) or (II) wherein
R3 is a C1-C4 alkyl group optionally having 1 to 5 fluorine atoms as a substituent(s).
- [5] The therapeutic agent according to any of [1] to [4], wherein the pyrimidine compound
is a compound represented by the formula (I) or (II) wherein
R5 is a phenyl group optionally having 1 or 2 substituents selected from the group consisting
of fluorine atoms and chlorine atoms.
- [6] The therapeutic agent according to any of [1] to [5], wherein the pyrimidine compound
is a compound represented by the formula (I) or (II) wherein
R1 is a methyl group, a tert-butyl group, or a cyclopropyl group.
- [7] The therapeutic agent according to any of [1] to [6], wherein the pyrimidine compound
is a compound represented by the formula (I) or (II) wherein
R2 is a methyl group, an ethyl group, a methoxymethyl group, or an ethoxymethyl group.
- [8] The therapeutic agent according to any of [1] to [7], wherein the pyrimidine compound
is a compound represented by the formula (I) or (II) wherein
R3 is a methyl group.
- [9] The therapeutic agent according to any of [1] to [8], wherein the pyrimidine compound
is a compound represented by the formula (I) or (II) wherein
R4 is a hydrogen atom.
- [10] The therapeutic agent according to any of [1] to [9], wherein the pyrimidine
compound is a compound represented by the formula (I) or (II) wherein
R5 is a phenyl group.
- [11] The therapeutic agent according to any of [1] to [10], wherein the pyrimidine
compound is a compound selected from the following (1) to (3):
- (1) 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-N-((R)-1-phenylethyl)-6-(prop-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide,
- (2) 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide,
and
- (3) 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(3,3-dimethylbut-1-yn-1-yl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide.
- [12] The therapeutic agent according to any of [1] to [11], wherein the pyrimidine
compound is 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide.
- [13] The therapeutic agent according to any of [1] to [12], wherein the disease associated
with EGFR is malignant tumor having EGFR overexpression, EGFR gene amplification,
or an EGFR mutation.
- [14] An EGFR inhibitor comprising a pyrimidine compound represented by the following
formula (I), or a pharmaceutically acceptable salt thereof as an active ingredient:
wherein R1 represents a C1-C4 alkyl group optionally having a C1-C4 alkoxy group as a substituent,
or a C3-C4 cycloalkyl group;
R2 represents a hydrogen atom, a halogen atom, a C1-C6 alkyl group optionally having
1 to 5 C1-C4 alkoxy groups or fluorine atoms each as a substituent(s), or a C1-C6
alkoxy group;
R3 represents a hydrogen atom, or a C1-C4 alkyl group optionally having 1 to 5 fluorine
atoms as a substituent(s);
R4 represents a hydrogen atom or a C1-C4 alkyl group; and
R5 represents a phenyl group optionally having 1 to 3 substituents selected from fluorine
atoms and chlorine atoms.
- [15] A therapeutic agent for EGFR-positive tumor, comprising a pyrimidine compound
represented by the following formula (I), or a pharmaceutically acceptable salt thereof
as an active ingredient:
wherein R1 represents a C1-C4 alkyl group optionally having a C1-C4 alkoxy group as a substituent,
or a C3-C4 cycloalkyl group;
R2 represents a hydrogen atom, a halogen atom, a C1-C6 alkyl group optionally having
1 to 5 C1-C4 alkoxy groups or fluorine atoms each as a substituent(s), or a C1-C6
alkoxy group;
R3 represents a hydrogen atom, or a C1-C4 alkyl group optionally having 1 to 5 fluorine
atoms as a substituent(s);
R4 represents a hydrogen atom or a C1-C4 alkyl group; and
R5 represents a phenyl group optionally having 1 to 3 substituents selected from fluorine
atoms and chlorine atoms.
- [16] A pharmaceutical composition for use in the treatment of a disease associated
with EGFR, the pharmaceutical composition comprising a pyrimidine compound represented
by the following formula (I), or a pharmaceutically acceptable salt thereof and a
pharmaceutically acceptable carrier:
wherein R1 represents a C1-C4 alkyl group optionally having a C1-C4 alkoxy group as a substituent,
or a C3-C4 cycloalkyl group;
R2 represents a hydrogen atom, a halogen atom, a C1-C6 alkyl group optionally having
1 to 5 C1-C4 alkoxy groups or fluorine atoms each as a substituent(s), or a C1-C6
alkoxy group;
R3 represents a hydrogen atom, or a C1-C4 alkyl group optionally having 1 to 5 fluorine
atoms as a substituent(s);
R4 represents a hydrogen atom or a C1-C4 alkyl group; and
R5 represents a phenyl group optionally having 1 to 3 substituents selected from fluorine
atoms and chlorine atoms.
- [17] The pharmaceutical composition according to [16], wherein the disease associated
with EGFR is malignant tumor having EGFR overexpression, EGFR gene amplification,
or an EGFR mutation.
[0013] The present invention also relates to the following aspects:
A compound represented by the formula (I) or a pharmaceutically acceptable salt thereof
for treating a disease associated with EGFR.
[0014] Use of a compound represented by the formula (I) or a pharmaceutically acceptable
salt thereof for treating a disease associated with EGFR.
[0015] Use of a compound represented by the formula (I) or a pharmaceutically acceptable
salt thereof for producing a therapeutic agent for a disease associated with EGFR.
[0016] A compound represented by the formula (I) or a pharmaceutically acceptable salt thereof
for treating EGFR-positive tumor.
[0017] Use of a compound represented by the formula (I) or a pharmaceutically acceptable
salt thereof for treating EGFR-positive tumor.
[0018] Use of a compound represented by the formula (I) or a pharmaceutically acceptable
salt thereof for producing a therapeutic agent for EGFR-positive tumor.
[0019] A method for treating a disease associated with EGFR, comprising administering an
effective amount of a compound represented by the formula (I) or a pharmaceutically
acceptable salt thereof to a subject in need thereof.
[0020] A method for treating EGFR-positive tumor, comprising administering an effective
amount of a compound represented by the formula (I) or a pharmaceutically acceptable
salt thereof to a subject in need thereof.
[0021] A compound represented by the formula (I) or a pharmaceutically acceptable salt thereof
for treating malignant tumor having exon 18 mutant EGFR.
[0022] Use of a compound represented by the formula (I) or a pharmaceutically acceptable
salt thereof for treating malignant tumor having exon 18 mutant EGFR.
[0023] Use of a compound represented by the formula (I) or a pharmaceutically acceptable
salt thereof for producing a therapeutic agent for malignant tumor having exon 18
mutant EGFR.
[0024] A method for treating malignant tumor having exon 18 mutant EGFR, comprising administering
an effective amount of a compound represented by the formula (I) or a pharmaceutically
acceptable salt thereof to a subject in need thereof.
[0025] A compound represented by the formula (I) or a pharmaceutically acceptable salt thereof
for treating malignant tumor having exon 19 mutant EGFR.
[0026] Use of a compound represented by the formula (I) or a pharmaceutically acceptable
salt thereof for treating malignant tumor having exon 19 mutant EGFR.
[0027] Use of a compound represented by the formula (I) or a pharmaceutically acceptable
salt thereof for producing a therapeutic agent for malignant tumor having exon 19
mutant EGFR.
[0028] A method for treating malignant tumor having exon 19 mutant EGFR, comprising administering
an effective amount of a compound represented by the formula (I) or a pharmaceutically
acceptable salt thereof to a subject in need thereof.
[0029] A compound represented by the formula (I) or a pharmaceutically acceptable salt thereof
for treating malignant tumor having exon 20 mutant EGFR.
[0030] Use of a compound represented by the formula (I) or a pharmaceutically acceptable
salt thereof for treating malignant tumor having exon 20 mutant EGFR.
[0031] Use of a compound represented by the formula (I) or a pharmaceutically acceptable
salt thereof for producing a therapeutic agent for malignant tumor having exon 20
mutant EGFR.
[0032] A method for treating malignant tumor having exon 20 mutant EGFR, comprising administering
an effective amount of a compound represented by the formula (I) or a pharmaceutically
acceptable salt thereof to a subject in need thereof.
[0033] A compound represented by the formula (I) or a pharmaceutically acceptable salt thereof
for treating malignant tumor having exon 21 mutant EGFR.
[0034] Use of a compound represented by the formula (I) or a pharmaceutically acceptable
salt thereof for treating malignant tumor having exon 21 mutant EGFR.
[0035] Use of a compound represented by the formula (I) or a pharmaceutically acceptable
salt thereof for producing a therapeutic agent for malignant tumor having exon 21
mutant EGFR.
[0036] A method for treating malignant tumor having exon 21 mutant EGFR, comprising administering
an effective amount of a compound represented by the formula (I) or a pharmaceutically
acceptable salt thereof to a subject in need thereof.
[0037] A compound represented by the formula (I) or a pharmaceutically acceptable salt thereof
for treating malignant tumor resistant to an existing EGFR inhibitor.
[0038] Use of a compound represented by the formula (I) or a pharmaceutically acceptable
salt thereof for treating malignant tumor resistant to an existing EGFR inhibitor.
[0039] Use of a compound represented by the formula (I) or a pharmaceutically acceptable
salt thereof for producing a therapeutic agent for malignant tumor resistant to an
existing EGFR inhibitor.
[0040] A method for treating malignant tumor resistant to an existing EGFR inhibitor, comprising
administering an effective amount of a compound represented by the formula (I) or
a pharmaceutically acceptable salt thereof to a subject in need thereof.
Advantageous Effects of Invention
[0041] The therapeutic agent for a disease associated with EGFR according to the present
invention provides a novel therapeutic method for treating a disease associated with
EGFR, or EGFR-positive tumor.
[0042] The compound of the present invention or a salt thereof has excellent inhibitory
activity against wild-type EGFR and mutant EGFR and has excellent brain penetration
properties, antitumor effect on malignant tumor, and effect of extending a survival
period.
Brief Description of Drawings
[0043]
[Figure 1] Figure 1 shows the antitumor effects of compounds of Example 2, 11 and
12 against models involving subcutaneous transplantation of the H1975-EGFRinsSVD cell
line.
[Figure 2] Figure 2 shows the body weight change percentage of mice when the compounds
of Examples 2, 11 and 12 were administered to the models involving subcutaneous transplantation
of the H1975-EGFRinsSVD cell line.
[Figure 3] Figure 3 shows the antitumor effects of the compound of Example 11 against
models involving direct brain transplantation of the Luciferase gene-introduced exon
20 insertion mutant EGFR expressing cell line (H1975-EGFRinsSVD-Luc).
[Figure 4] Figure 4 shows the survival rate of mice when the compound of Example 11
was administered to the models involving direct brain transplantation of the Luciferase
gene-introduced exon 20 insertion mutant EGFR expressing cell line (H1975-EGFRinsSVD-Luc).
Description of Embodiments
[0044] One embodiment of the present invention relates to a compound represented by the
following formula (I), or a salt thereof:
(wherein R
1 to R
5 represents as defined above.)
[0045] One preferred embodiment of the present invention is a pyrimidine compound represented
by the following formula (II), or a salt thereof
(wherein R
1 to R
5 represents as defined above.)
[0046] The compound represented by the above formula (I) or formula (II) of the present
invention is a compound having pyrrolo[2,3-d]pyrimidine as a basic structure, and
this is a novel compound described in none of the aforementioned prior art publications,
etc.
[0047] In the present description, specific examples of the "halogen atom" may include
a chlorine atom, a bromine atom, a fluorine atom, and an iodine atom. Among these,
a chlorine atom and a fluorine atom are preferable, and a fluorine atom is more preferable.
[0048] In the present description, the "alkyl group" means a linear or branched saturated
hydrocarbon group. Specific examples of the alkyl group may include a methyl group,
an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl
group, a sec-butyl group, a tert-butyl group, a pentyl group, and a hexyl group. Among
these, a linear or branched alkyl group containing 1 to 4 carbon atoms is preferable,
and a methyl group and a tert-butyl group are more preferable.
[0049] In the present description, the "haloalkyl group" means a linear or branched saturated
hydrocarbon group, in which one to all hydrogen atoms are substituted with the above-described
halogen atoms. Specific examples of the haloalkyl group may include a monofluoromethyl
group, a difluoromethyl group, a trifluoromethyl group, a 1-fluoroethyl group, a 2-fluoroethyl
group, a 1,1-difluoroethyl group, a 1,2-difluoroethyl group, a 2,2-difluoroethyl group,
a 2,2,2-trifluoroethyl group, a monochloromethyl group, a dichloromethyl group, a
trichloromethyl group, a 1-chloroethyl group, a 2-chloroethyl group, and a 1,1-dichloroethyl
group. Among these, a linear or branched saturated hydrocarbon group containing 1
to 6 carbon atoms, in which 1 to 3 hydrogen atoms are substituted with the above-described
halogen atoms, is preferable, and a monofluoromethyl group is more preferable.
[0050] In the present description, the "cycloalkyl group" means a monocyclic or polycyclic
saturated hydrocarbon group containing 3 to 7 carbon atoms. Specific examples of the
cycloalkyl group may include a cyclopropyl group, a cyclobutyl group, a cyclopentyl
group, a cyclohexyl group, and a cycloheptyl group. Among these, a cyclopropyl group
and a cyclobutyl group are preferable.
[0051] In the present description, the "aromatic hydrocarbon group" means a cyclic substituent
consisting of carbon and hydrogen, having an unsaturated bond, in which 4e + 2 (wherein
e represents an integer of 1 or greater) electrons are contained in the cyclic π electron
system.
[0052] In the present description, the "C6-C14 aromatic hydrocarbon group" means a monocyclic
or polycyclic aromatic hydrocarbon group containing 6 to 14 carbon atoms. Specific
examples of the C6-C14 aromatic hydrocarbon group may include a phenyl group, a naphthyl
group, a tetrahydronaphthyl group, and an anthracenyl group. Among these, a phenyl
group is preferable.
[0053] In the present description, the "aralkyl group" means the above-described alkyl group
substituted with the above-described aromatic hydrocarbon group. Specific examples
of the aralkyl group may include C7-C16 aralkyl groups such as a benzyl group, a phenylethyl
group, a phenylpropyl group, a naphthylmethyl group, and a naphthylethyl group. Among
these, a benzyl group is preferable.
[0054] In the present description, the "unsaturated hydrocarbon group" means a linear or
branched hydrocarbon group containing 2 to 6 carbon atoms, which comprises at least
one carbon-carbon double bond or triple bond. Specific examples of the unsaturated
hydrocarbon group may include a vinyl group, an allyl group, a methylvinyl group,
a propenyl group, a butenyl group, a pentenyl group, a hexenyl group, an ethynyl group,
and a 2-propynyl group. Among these, a vinyl group, an allyl group, and a 1-propenyl
group are preferable.
[0055] In the present description, the "alkenyl group" means a linear or branched hydrocarbon
group containing 2 to 6 carbon atoms, which comprises at least one carbon-carbon double
bond. Specific examples of the alkenyl group may include C2-C6 alkenyl groups, such
as a vinyl group, an allyl group, a 2-methyl-2-propenyl group, an isopropenyl group,
a 1-, 2- or 3-butenyl group, a 2-, 3- or 4-pentenyl group, a 2-methyl-2-butenyl group,
a 3-methyl-2-butenyl group, and a 5-hexenyl group. Among these, a vinyl group, an
allyl group, a 1-propenyl group, and a 2-methyl-2-propenyl group are preferable.
[0056] In the present description, the "alkynyl group" means a linear or branched unsaturated
hydrocarbon group having at least one triple bond (for example, 1 or 2, and preferably
1 triple bond). Specific examples of the alkynyl group may include C2-C6 alkynyl groups
such as an ethynyl group, a 1- or 2-propynyl group, a 1-, 2- or 3-butynyl group, and
a 1-methyl-2-propynyl group. Among these, an ethynyl group and a 2-propynyl group
are preferable.
[0057] In the present description, the "C3-C10 cyclic unsaturated hydrocarbon group" means
a monocyclic or polycyclic hydrocarbon group containing 3 to 10 carbon atoms, which
comprises at least one carbon-carbon double bond. Specific examples of the C3-C10
cyclic unsaturated hydrocarbon group may include a cyclopropenyl group, a cyclobutenyl
group, a cyclopentenyl group, a cyclohexenyl group, a cycloheptenyl group, a cyclooctenyl
group, and a cyclononyl group. Among these, a monocyclic or polycyclic hydrocarbon
group containing 3 to 7 carbon atoms, which comprises at least one carbon-carbon double
bond, is preferable, and a cyclopropenyl group is more preferable.
[0058] In the present description, the "alkoxy group" means an oxy group having the above-described
alkyl group. Specific examples of the alkoxy group may include C1-C6 alkoxy groups
such as a methoxy group, an ethoxy group, an n-propoxy group, an isopropoxy group,
an n-butoxy group, an isobutoxy group, a sec-butoxy group, a tert-butoxy group, a
pentyloxy group, an isopentyloxy group, and a hexyloxy group. Among these, a methoxy
group and an ethoxy group are preferable, and a methoxy group is more preferable.
[0059] In the present description, the "haloalkoxy group" may include the above-described
alkoxy group having at least one halogen atom (preferably 1 to 13, and more preferably
1 to 3 halogen atoms). Specific examples of the haloalkoxy group may include C1-C6
haloalkoxy groups such as a fluoromethoxy group, a difluoromethoxy group, a trifluoromethoxy
group, a trichloromethoxy group, a fluoroethoxy group, a 1,1,1-trifluoroethoxy group,
a monofluoro-n-propoxy group, a perfluoro-n-propoxy group, and a perfluoro-isopropoxy
group.
[0060] In the present description, the "cycloalkoxy group" means an oxy group having the
above-described cycloalkyl group. Specific examples of the cycloalkoxy group may include
C3-C7 cycloalkoxy groups such as a cyclopropoxy group, a cyclobutoxy group, a cyclopentyloxy
group, a cyclohexyloxy group, and a cycloheptyloxy group. Among these, a cyclobutoxy
group, a cyclopentyloxy group, and a cyclohexyloxy group are preferable.
[0061] In the present description, the "aralkyloxy group" means an oxy group having the
above-described aralkyl group. Specific examples of the aralkyloxy group may include
C7-C20 aralkyloxy groups such as a benzyloxy group, a phenethyloxy group, a naphthylmethyloxy
group, and a fluorenylmethyloxy group. Among these, a benzyloxy group is preferable.
[0062] In the present description, the "alkylthio group" means a thioxy group having the
above-described alkyl group. Specific examples of the alkylthio group may include
C1-C6 alkylthio groups such as a methylthio group, an ethylthio group, an n-propylthio
group, an isopropylthio group, an n-butylthio group, an isobutylthio group, a tert-butylthio
group, an n-pentylthio group, an isopentylthio group, and a hexylthio group. Among
these, a methylthio group, an ethylthio group, and an n-propylthio group are preferable.
[0063] In the present description, the "alkoxyalkyl group" means the above-described alkyl
group having at least one of the above-described alkoxy groups. Specific examples
of the alkoxyalkyl group may include C1-C6 alkoxy-C1-C6 alkyl groups such as a methoxymethyl
group, an ethoxyethyl group, a methoxyethyl group, and a methoxypropyl group.
[0064] In the present description, the "alkylamino group" means an amino group in which
1 or 2 hydrogen atoms are substituted with a linear or branched hydrocarbon group(s)
containing 1 to 6 carbon atoms. Specific examples of the alkylamino group may include
a methylamino group, an ethylamino group, a dimethylamino group, a diethylamino group,
and an ethylmethylamino group. Among these, preferable is an amino group in which
1 or 2 hydrogen atoms are substituted with a linear or branched hydrocarbon group
containing 1 to 3 carbon atoms.
[0065] In the present description, the "monoalkylamino group" means an amino group in which
one hydrogen atom is substituted with a linear or branched hydrocarbon group. Specific
examples of the monoalkylamino group may include a methylamino group, an ethylamino
group, an n-propylamino group, an isopropylamino group, an n-butylamino group, an
isobutylamino group, a sec-butylamino group, a tert-butylamino group, a pentylamino
group, and a hexylamino group. Among these, preferable is an amino group in which
one hydrogen atom is substituted with a linear or branched hydrocarbon group containing
1 to 3 carbon atoms.
[0066] In the present description, the "dialkylamino group" means an amino group in which
two hydrogen atoms are substituted with linear or branched hydrocarbon groups containing
1 to 6 carbon atoms. Specific examples of the dialkylamino group may include a dimethylamino
group, a diethylamino group, and an ethylmethylamino group. Among these, an amino
group in which two hydrogen atoms are substituted with linear or branched hydrocarbon
groups containing 1 to 3 carbon atoms is preferable, and a dimethylamino group is
more preferable.
[0067] In the present description, the "acyl group" means a formyl group in which a hydrogen
atom is substituted with a linear or branched hydrocarbon group. Specific examples
of the acyl group may include an acetyl group, an n-propanoyl group, an isopropanoyl
group, an n-butyloyl group, and a tert-butyloyl group. Among these, a formyl group
in which a hydrogen atom is substituted with a linear or branched hydrocarbon group
containing 1 to 3 carbon atoms is preferable, and an acetyl group is more preferable.
[0068] In the present description, the "acyloxy group" means an oxy group having the above-described
acyl group. Specific examples of the acyloxy group may include an alkylcarbonyloxy
group and an arylcarbonyloxy group. Among these, an oxy group in which a hydrogen
atom of formyl group is substituted with a linear or branched hydrocarbon group containing
1 to 3 carbon atoms is preferable, and an alkylcarbonyloxy group is more preferable.
[0069] In the present description, the "alkoxycarbonyl group" means a carbonyl group having
the above-described alkoxy group. Specific examples of the alkoxycarbonyl group may
include (C1-C6alkoxy)carbonyl groups such as a methoxycarbonyl group, an ethoxycarbonyl
group, a propoxycarbonyl group, an isopropoxycarbonyl group, a butoxycarbonyl group,
an isobutoxycarbonyl group, a tert-butoxycarbonyl group, a pentyloxycarbonyl group,
an isopentyloxycarbonyl group, and a hexyloxycarbonyl group. Among these, a tert-butoxycarbonyl
group is preferable.
[0070] In the present description, the "aralkyloxycarbonyl group" means a carbonyl group
having the above-described aralkyloxy. Specific examples of the aralkyloxycarbonyl
group may include (C6-C20 aralkyl)oxycarbonyl groups such as a benzyloxycarbonyl group,
a phenethyloxycarbonyl group, a naphthylmethyloxycarbonyl group, and a fluorenylmethyloxycarbonyl
group. Among these, a benzyloxycarbonyl group is preferable.
[0071] In the present description, the "saturated heterocyclic group" means a monocyclic
or polycyclic saturated heterocyclic group having at least one heteroatom (preferably
1 to 5, and more preferably 1 to 3 heteroatoms) selected from nitrogen atoms, oxygen
atoms, and sulfur atoms. Specific examples of the saturated heterocyclic group may
include an aziridinyl group, an azetidinyl group, an imidazolidinyl group, a morpholino
group, a pyrrolidinyl group, a piperidinyl group, a piperazinyl group, a tetrahydrofuranyl
group, a tetrahydropyranyl group, a tetrahydrothiophenyl group, a thiazolidinyl group,
and an oxazolidinyl group. Among these, an azetidinyl group, a pyrrolidinyl group,
and a piperidinyl group are preferable, and an azetidinyl group and a pyrrolidinyl
group are more preferable.
[0072] In the present description, the "unsaturated heterocyclic group" means a monocyclic
or polycyclic completely unsaturated or partially unsaturated heterocyclic group having
at least one heteroatom (preferably 1 to 5, and more preferably 1 to 3 heteroatoms)
selected from nitrogen atoms, oxygen atoms, and sulfur atoms. Specific examples of
the unsaturated heterocyclic group may include an imidazolyl group, a thienyl group,
a pyrrolyl group, an oxazolyl group, an isoxazolyl group, a thiazolyl group, an isothiazolyl
group, a thiadiazolyl group, an oxadiazolyl group, a pyrazolyl group, a triazolyl
group, a tetrazolyl group, a pyridyl group, a pyrazyl group, a pyrimidinyl group,
a pyridazinyl group, an indolyl group, an isoindolyl group, an indazolyl group, a
triazolopyridyl group, a benzimidazolyl group, a benzoxazolyl group, a benzothiazolyl
group, a benzothienyl group, a furanyl group, a benzofuranyl group, a purinyl group,
a quinolyl group, an isoquinolyl group, a quinazolinyl group, a quinoxalyl group,
a methylenedioxyphenyl group, an ethylenedioxyphenyl group, and a dihydrobenzofuranyl
group. Among these, an imidazolyl group, a pyrazolyl group, a thiazolyl group, an
isoxazolyl group, and a furanyl group are preferable; an imidazolyl group, a pyrazolyl
group, and a thiazolyl group are more preferable; and an imidazolyl group is most
preferable.
[0073] In the present description, the "saturated heterocyclic oxy group" means an oxy group
having the above-described saturated heterocyclic group. Specific examples of the
saturated heterocyclic oxy group may include a morpholinyloxy group, a 1-pyrrolidinyloxy
group, a piperidinooxy group, a piperazinyloxy group, a 4-methyl-1-piperazinyloxy
group, a tetrahydrofuranyloxy group, a tetrahydropyranyloxy group, a tetrahydrothiophenyloxy
group, a thiazolidinyloxy group, and an oxazolidinyloxy group. Among these, a 1-pyrrolidinyloxy
group, a piperidinooxy group, and a piperazinyloxy group are preferable.
[0074] In the compound represented by the formula (I) or the formula (II) of the present
invention, R
1 is a C1-C4 alkyl group optionally having a C1-C4 alkoxy group as a substituent, or
a C3-C4 cycloalkyl group.
[0075] The "C1-C4 alkoxy group" in the "C1-C4 alkyl group optionally having a C1-C4 alkoxy
group as a substituent" represented by R
1 is preferably a methoxy group or an ethoxy group, and most preferably a methoxy group.
Herein, the number of substituents is preferably 1 to 3, and most preferably 1. When
the C1-C4 alkyl group has two or more substituents, the substituents may be identical
to or different from each other.
[0076] The "C1-C4 alkyl group" in the "C1-C4 alkyl group optionally having a C1-C4 alkoxy
group as a substituent" represented by R
1 is preferably a methyl group, an ethyl group, an n-propyl group, an isopropyl group,
or a tert-butyl group, more preferably a methyl group, an ethyl group, an isopropyl
group, or a tert-butyl group, and most preferably a methyl group or a tert-butyl group.
[0077] The "C1-C4 alkyl group optionally having a C1-C4 alkoxy group as a substituent" represented
by R
1 is preferably a C1-C4 alkyl group having 1 to 3 methoxy groups as substituents, more
preferably a methyl group, an ethyl group, an isopropyl group, a tert-butyl group,
or a 1-methyl-1-methoxyethyl group, and most preferably a methyl group or a tert-butyl
group.
[0078] The "C3-C4 cycloalkyl group" represented by R
1 is preferably a cyclopropyl group or a cyclobutyl group, and most preferably a cyclopropyl
group.
[0079] R
1 is preferably a C1-C4 alkyl group optionally having 1 to 3 C1-C4 alkoxy groups as
substituents, or a C3-C4 cycloalkyl group.
[0080] R
1 is more preferably a C1-C4 alkyl group optionally having 1 to 3 methoxy groups as
substituents, or a C3-C4 cycloalkyl group.
[0081] R
1 is further preferably a methyl group, an ethyl group, an isopropyl group, a tert-butyl
group, a 1-methyl-1-methoxyethyl group, or a cyclopropyl group.
[0082] R
1 is most preferably a methyl group, a tert-butyl group, or a cyclopropyl group.
[0083] In the compound represented by the formula (I) or the formula (II) of the present
invention, R
2 is a hydrogen atom, a halogen atom, a C1-C6 alkyl group optionally having 1 to 5
C1-C4 alkoxy groups or fluorine atoms each as a substituent(s), or a C1-C6 alkoxy
group.
[0084] The "halogen atom" represented by R
2 is preferably a fluorine atom or a chlorine atom.
[0085] The "C1-C4 alkoxy group" in the "C1-C6 alkyl group optionally having 1 to 5 C1-C4
alkoxy groups or fluorine atoms each as a substituent(s)" represented by R
2 is preferably a methoxy group or an ethoxy group, and most preferably a methoxy group.
[0086] The "C1-C6 alkyl group optionally having 1 to 5 C1-C4 alkoxy groups or fluorine atoms
each as a substituent(s)" represented by R
2 is preferably a methyl group, an ethyl group, an n-propyl group, an isopropyl group,
or a tert-butyl group, and most preferably a methyl group.
[0087] The "C1-C6 alkyl group" in the "C1-C6 alkyl group optionally having 1 to 5 C1-C4
alkoxy groups or fluorine atoms each as a substituent(s)" represented by R
2 is preferably a C1-C6 alkyl group optionally having 1 to 5 methoxy groups, ethoxy
groups, or fluorine atoms as a substituent(s) (specifically, a methyl group, a methoxymethyl
group, an ethoxymethyl group, a methoxyethyl group, an ethoxyethyl group, a fluoromethyl
group, a difluoromethyl group, a trifluoromethyl group, etc.), more preferably a C1-C6
alkyl group, further preferably a methyl group, an ethyl group, an n-propyl group,
an isopropyl group, or a tert-butyl group, and most preferably a methyl group.
[0088] The "C1-C6 alkoxy group" represented by R
2 is preferably a methoxy group or an ethoxy group, and most preferably a methoxy group.
[0089] R
2 is preferably a C1-C6 alkyl group optionally having 1 to 5 C1-C4 alkoxy groups or
fluorine atoms each as a substituent(s). In one embodiment, R
2 is a C1-C6 alkyl group optionally having 1 to 5 methoxy groups, ethoxy groups, or
fluorine atoms as a substituent(s). In another embodiment, R
2 is a methyl group, an ethyl group, an n-propyl group, an isopropyl group, or a tert-butyl
group (preferably a methyl group or an ethyl group, and more preferably a methyl group),
each optionally having 1 to 5 methoxy groups, ethoxy groups, or fluorine atoms as
a substituent(s).
[0090] R
2 is more preferably a C1-C6 alkyl group optionally having 1 to 5 C1-C4 alkoxy groups
as a substituent(s). In one embodiment, R
2 is a C1-C6 alkyl group optionally having 1 to 5 methoxy groups or ethoxy groups as
a substituent(s). In another embodiment, R
2 is a methyl group, an ethyl group, an n-propyl group, an isopropyl group, or a tert-butyl
group (preferably a methyl group or an ethyl group, and more preferably a methyl group)
each optionally having 1 to 5 methoxy groups or ethoxy groups as a substituent(s).
In a further embodiment, R
2 is a methyl group, an ethyl group, a methoxymethyl group, or an ethoxymethyl group.
[0091] R
2 is even more preferably a C1-C6 alkyl group.
[0092] R
2 is further preferably a methyl group, an ethyl group, an n-propyl group, an isopropyl
group, or a tert-butyl group.
[0093] R
2 is particularly preferably a methyl group or an ethyl group.
[0094] R
2 is most preferably a methyl group.
[0095] In the compound represented by the formula (I) or the formula (II) of the present
invention, R
3 is a hydrogen atom, or a C1-C4 alkyl group optionally having 1 to 5 fluorine atoms
as a substituent(s).
[0096] The "C1-C4 alkyl group" in the "C1-C4 alkyl group optionally having 1 to 5 fluorine
atoms as a substituent(s)" represented by R
3 is preferably a methyl group, an ethyl group, an n-propyl group, an isopropyl group,
or a tert-butyl group, more preferably a methyl group or an ethyl group, and most
preferably a methyl group.
[0097] The "C1-C4 alkyl group optionally having 1 to 5 fluorine atoms as a substituent(s)"
represented by R
3 is preferably a methyl group, a fluoromethyl group, a difluoromethyl group, a trifluoromethyl
group, or an ethyl group, more preferably a methyl group, a trifluoromethyl group,
or an ethyl group, and most preferably a methyl group.
[0098] R
3 is preferably a C1-C4 alkyl group optionally having 1 to 5 fluorine atoms as a substituent(s).
[0099] R
3 is more preferably a methyl group, a fluoromethyl group, a difluoromethyl group,
a trifluoromethyl group, an ethyl group, a fluoroethyl group, a difluoroethyl group,
a trifluoroethyl group, an n-propyl group, an isopropyl group, or a tert-butyl group.
[0100] R
3 is even more preferably a methyl group, a fluoromethyl group, a difluoromethyl group,
a trifluoromethyl group, or an ethyl group.
[0101] R
3 is further preferably a methyl group, a trifluoromethyl group, or an ethyl group.
[0102] R
3 is particularly preferably a methyl group or an ethyl group.
[0103] R
3 is most preferably a methyl group.
[0104] In the compound represented by the formula (I) or the formula (II) of the present
invention, R
4 is a hydrogen atom or a C1-C4 alkyl group.
[0105] The "C1-C4 alkyl group" represented by R
4 is preferably a methyl group, an ethyl group, an n-propyl group, an isopropyl group,
or a tert-butyl group, more preferably a methyl group or an ethyl group, and most
preferably a methyl group.
[0106] R
4 is preferably a hydrogen atom, a methyl group, an ethyl group, an n-propyl group,
an isopropyl group, or a tert-butyl group.
[0107] R
4 is more preferably a hydrogen atom, a methyl group, or an ethyl group.
[0108] R
4 is further preferably a hydrogen atom or a methyl group.
[0109] R
4 is most preferably a hydrogen atom.
[0110] In the compound represented by the formula (I) or the formula (II) of the present
invention, R
5 is a phenyl group optionally having 1 to 3 substituents selected from the group consisting
of fluorine atoms and chlorine atoms.
[0111] R
5 is preferably a phenyl group optionally having 1 or 2 substituents selected from
the group consisting of fluorine atoms and chlorine atoms.
[0112] R
5 is more preferably a phenyl group, a 2-fluorophenyl group, a 3-chlorophenyl group,
a 2,3-difluorophenyl group, a 2,4-difluorophenyl group, or a 3,5-difluorophenyl group.
[0113] R
5 is most preferably a phenyl group.
[0114] The compound of the present invention is preferably the compound represented by
the formula (I) or the formula (II), or a salt thereof, wherein, in the formula (I)
or the formula (II),
R1 is a C1-C4 alkyl group optionally having a C1-C4 alkoxy group as a substituent, or
a C3-C4 cycloalkyl group,
R2 is a C1-C6 alkyl group optionally having 1 to 5 C1-C4 alkoxy groups as a substituent(s),
R3 is a C1-C4 alkyl group optionally having 1 to 5 fluorine atoms as a substituent(s),
R4 is a hydrogen atom or a C1-C4 alkyl group, and
R5 is a phenyl group optionally having 1 or 2 substituents selected from the group consisting
of fluorine atoms and chlorine atoms.
[0115] The compound of the present invention is more preferably the compound represented
by the formula (I) or the formula (II), or a salt thereof, wherein, in the formula
(I) or the formula (II),
R1 is a methyl group, an ethyl group, an n-propyl group, an isopropyl group, a tert-butyl
group, a 1-methyl-1-methoxyethyl group, a cyclopropyl group, or a cyclobutyl group,
R2 is a methyl group, an ethyl group, an n-propyl group, a tert-butyl group, a methoxymethyl
group, or an ethoxymethyl group,
R3 is a methyl group, a fluoromethyl group, a difluoromethyl group, a trifluoromethyl
group, an ethyl group, a fluoroethyl group, a difluoroethyl group, a trifluoroethyl
group, an n-propyl group, an isopropyl group, or a tert-butyl group,
R4 is a hydrogen atom, a methyl group, an ethyl group, an n-propyl group, an isopropyl
group, or a tert-butyl group, and
R5 is a phenyl group, a 2-fluorophenyl group, a 3-fluorophenyl group, a 2,4-difluorophenyl
group, a 2,3-difluorophenyl group, a 3,5-difluorophenyl group, a 2-chlorophenyl group,
a 3-chlorophenyl group, a 2,4-dichlorophenyl group, or a 3,5-dichlorophenyl group.
[0116] The compound of the present invention is even more preferably the compound represented
by the formula (II), or a salt thereof, wherein, in the formula (II),
R1 is a methyl group, an ethyl group, an isopropyl group, a tert-butyl group, a 1-methyl-1-methoxyethyl
group, or a cyclopropyl group,
R2 is a methyl group, an ethyl group, a methoxymethyl group, or an ethoxymethyl group,
R3 is a methyl group, a fluoromethyl group, a difluoromethyl group, a trifluoromethyl
group, or an ethyl group,
R4 is a hydrogen atom, a methyl group, or an ethyl group, and
R5 is a phenyl group, a 2-fluorophenyl group, a 3-chlorophenyl group, a 2,3-difluorophenyl
group, a 2,4-difluorophenyl group, or a 3,5-difluorophenyl group.
[0117] The compound of the present invention is further preferably the compound represented
by the formula (II), or a salt thereof, wherein, in the formula (II),
R1 is a methyl group, a tert-butyl group, or a cyclopropyl group,
R2 is a methyl group, an ethyl group, a methoxymethyl group, or an ethoxymethyl group,
R3 is a methyl group, a trifluoromethyl group, or an ethyl group,
R4 is a hydrogen atom or a methyl group, and
R5 is a phenyl group.
[0118] The compound of the present invention is further preferably the compound represented
by the formula (II), or a salt thereof, wherein, in the formula (II),
R1 is a methyl group, a tert-butyl group, or a cyclopropyl group,
R2 is a methyl group, an ethyl group, or a methoxymethyl group,
R3 is a methyl group,
R4 is a hydrogen atom, and
R5 is a phenyl group.
[0119] The compound of the present invention is particularly preferably the compound represented
by the formula (II), or a salt thereof, wherein, in the formula (II),
R1 is a methyl group, a tert-butyl group, or a cyclopropyl group,
R2 is a methyl group,
R3 is a methyl group,
R4 is a hydrogen atom, and
R5 is a phenyl group.
[0120] Specific examples of the compound represented by the formula (I) or the formula (II)
may include compounds produced in the following Examples, but are not limited thereto.
[0121] One embodiment of the present invention relates to a compound selected from the following
(1) to (18), or a salt thereof. One embodiment of the present invention relates to
a compound selected from the following (1) to (15), or a salt thereof.
- (1) 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-N-((R)-1-phenylethyl)-6-(prop-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide,
- (2) 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide,
- (3) 7-((3R,SS)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(3,3-dimethylbut-1-yn-1-yl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide,
- (4) 7-(R)-((3R,SS)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-N-((R)-1-(3,5-difluorophenyl)ethyl)-6-(prop-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide,
- (5) 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-N-(2-phenylpropan-2-yl)-6-(prop-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide,
- (6) 7-((3R,SS)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-N-((R)-1-phenylpropyl)-6-(prop-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide,
- (7) 7-((3R,SS)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-N-(2-(2-fluorophenyl)propan-2-yl)-6-(prop-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide,
- (8) 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-N-((R)-1-(3-chlorophenyl)ethyl)-6-(prop-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide,
- (9) 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-N-((R)-1-(2,4-difluorophenyl)ethyl)-6-(prop-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide,
- (10) 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(prop-1-yn-1-yl)-N-((S)-2,2,2-trifluoro-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide,
- (11) 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-(2-phenylpropan-2-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide,
- (12) 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-(2,3-difluorophenyl)ethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide,
- (13) 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(3-methoxy-3-methylbut-1-yn-1-yl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide,
- (14) 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(but-1-yn-1-yl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide,
- (15) 7-((3R,SS)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-N-(2-(2-fluorophenyl)propan-2-yl)-6-(3-methylbut-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide,
- (16) 7-((3R,SS)-1-acryloyl-5-ethylpyrrolidin-3-yl)-4-amino-N-((R)-1-phenylethyl)-6-(prop-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide,
- (17) 7-((3R,5S)-1-acryloyl-5-ethylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide,
- (18) 7-((3R,5R)-1-acryloyl-5-(methoxymethyl)pyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide,
and
- (19) 7-((3R,5R)-1-acryloyl-5-(ethoxymethyl)pyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide.
[0122] A preferred example of the compound represented by the formula (I) or the formula
(II) may be a compound selected from the following (1) to (3), or a salt thereof.
- (1) 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-N-((R)-1-phenylethyl)-6-(prop-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide,
- (2) 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide,
and
- (3) 7-((3R,SS)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(3,3-dimethylbut-1-yn-1-yl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide.
[0123] The most preferable pyrimidine compound of the present invention is 7-((3R,5 S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide.
< Method for producing compound represented by formula (I) >
[0124] The compound according to the present invention can be produced by, for example,
the following production method or the methods described in the Examples. However,
the method for producing the compound according to the present invention is not limited
to these examples.
[0125] The compounds (I) and (II) of the present invention can be produced by applying,
for example, the following production method.
< Production Method >
[0126]
[0127] In the above process, L
1, L
2, and L
3, which are the same or different, each represent a leaving group; P
1 and P
2, which are the same or different, each represent a protective group; and other symbols
are as defined above.
< Step 1 >
[0128] This step is a method of obtaining a compound represented by the formula 3 by performing
a Mitsunobu reaction between a compound represented by the formula 1 and a compound
represented by the formula 2 that is a commercially available compound or can be produced
by a known method. The Mitsunobu reaction is generally carried out in the presence
of a Mitsunobu reagent and a phosphine reagent.
[0129] The compound represented by the formula 2 (in the formula 2, P
1 represents a protective group for an amino group) can be used in an amount of 1 to
10 equivalents, and preferably 1 to 3 equivalents, based on the amount of the compound
represented by the formula 1 (1 mole).
[0130] The "protective group for an amino group" is not particularly limited, as long as
it has a protective function. Examples of the protective group for an amino group
may include: aralkyl groups, such as a benzyl group, a p-methoxybenzyl group, a 3,4-dimethoxybenzyl
group, an o-nitrobenzyl group, a p-nitrobenzyl group, a benzhydryl group, a trityl
group, and a cumyl group; lower alkanoyl groups, such as, for example, a formyl group,
an acetyl group, a propionyl group, a butyryl group, a pivaloyl group, a trifluoroacetyl
group, and a trichloroacetyl group; for example, benzoyl groups; arylalkanoyl groups,
such as, for example, a phenylacetyl group and a phenoxyacetyl group; lower alkoxycarbonyl
groups, such as, for example, a methoxycarbonyl group, an ethoxycarbonyl group, a
propyloxycarbonyl group, and a tert-butoxycarbonyl group; aralkyloxycarbonyl groups,
such as, for example, a p-nitrobenzyloxycarbonyl group and a phenethyloxycarbonyl
group; lower alkylsilyl groups, such as, for example, a trimethylsilyl group and a
tert-butyldimethylsilyl group; for example, tetrahydropyranyl groups; for example,
trimethylsilylethoxymethyl groups; lower alkylsulfonyl groups, etc., such as, for
example, a methylsulfonyl group, an ethylsulfonyl group, and a tert-butylsulfonyl
group; lower alkylsulfinyl groups, etc., such as for example, a tert-butylsulfinyl
group; arylsulfonyl groups, etc., such as, for example, a benzenesulfonyl group and
a toluenesulfonyl group; and imide groups, such as, for example, a phthalimide group.
Among these, a trifluoroacetyl group, an acetyl group, a tert-butoxycarbonyl group,
a benzyloxycarbonyl group, a trimethylsilylethoxymethyl group, or a cumyl group is
particularly preferable.
[0131] As a Mitsunobu reagent, diethyl azodicarboxylate, diisopropyl azodicarboxylate or
the like is used. Such a Mitsunobu reagent is used in an amount of generally approximately
1 to 100 moles, and preferably approximately 1 to 10 moles, based on the compound
represented by the formula 1 (1 mole).
[0132] As a phosphine reagent, triphenylphosphine, tributylphosphine, trifurylphosphine
or the like is used. Such a phosphine reagent is used in an amount of generally approximately
1 to 100 moles, and preferably approximately 1 to 10 moles, based on the compound
represented by the formula 1 (1 mole).
[0133] The solvent is not particularly limited, as long as it does not affect the reaction.
Examples of the solvent may include hydrocarbons (e.g., benzene, toluene, xylene,
etc.), halogenated hydrocarbons (e.g., chloroform, 1,2-dichloroethane, etc.), nitriles
(e.g., acetonitrile, etc.), ethers (e.g., dimethoxyethane, tetrahydrofuran, etc.),
alcohols (e.g., methanol, ethanol, etc.), aprotic polar solvents (e.g., N,N-dimethylformamide,
dimethyl sulfoxide, hexamethylphosphoramide, etc.), water, and mixtures thereof. The
reaction time is 0.1 to 100 hours, and preferably 0.5 to 24 hours. Thereafter, the
reaction temperature is 0°C to the temperature at which the solvent is boiled, and
preferably 0°C to 100° C.
[0134] The thus obtained compound represented by the formula 3 can be isolated and purified
by known separation and purification means, or can be subjected to the subsequent
step without isolation and purification.
< Step 2 >
[0135] This step is a method of obtaining a compound represented by the formula 4 by allowing
the compound represented by the formula 3 to react with ammonia or a salt thereof.
[0136] The ammonia or a salt thereof can be used in an amount of 1 to 1000 equivalents,
and preferably 1 to 100 equivalents, based on the amount of the compound represented
by the formula 3 (1 mole).
[0137] The solvent is not particularly limited, as long as it does not affect the reaction.
Examples of the solvent may include hydrocarbons (e.g., benzene, toluene, xylene,
etc.), halogenated hydrocarbons (e.g., chloroform, 1,2-dichloroethane, etc.), nitriles
(e.g., acetonitrile, etc.), ethers (e.g., dimethoxyethane, tetrahydrofuran, etc.),
alcohols (e.g., methanol, ethanol, etc.), aprotic polar solvents (e.g., N,N-dimethylformamide,
dimethyl sulfoxide, hexamethylphosphoramide, etc.), water, and mixtures thereof. The
reaction time is 0.1 to 100 hours, and preferably 0.5 to 24 hours. Thereafter, the
reaction temperature is 0°C to the temperature at which the solvent is boiled, and
preferably 0°C to 150° C.
[0138] The thus obtained compound represented by the formula 4 can be isolated and purified
by known separation and purification means, or can be subjected to the subsequent
step without isolation and purification.
< Step 3 >
[0139] This step is a method of obtaining a compound represented by the formula 5 by reacting
the compound represented by the formula 4 under a carbon monoxide atmosphere, for
example, in the presence of a transition metal catalyst, a base and alcohol.
[0140] In this step, the pressure of the carbon monoxide is generally from 1 to 20 atmospheres,
and preferably 1 to 10 atmospheres.
[0141] Examples of the alcohol may include methanol, ethanol, propanol, isopropanol, diethylaminoethanol,
isobutanol, 4-(2-hydroxyethyl)morpholine, 3-morpholinopropanol, and diethylaminopropanol.
[0142] The alcohol is used in an amount of generally approximately 1 to 100 moles, and preferably
approximately 1 to 50 moles, based on the amount of the compound represented by the
formula 4 (1 mole).
[0143] Examples of the transition metal catalyst used herein may include palladium catalysts
(e.g., palladium acetate, palladium chloride, tetrakistriphenylphosphine palladium,
palladium carbon, etc.). A ligand (e.g., triphenylphosphine, tri-tert-butylphosphine,
etc.) may be added, as necessary. The amount of the transition metal catalyst used
is different depending on the type of the catalyst. The transition metal catalyst
is used in an amount of generally approximately 0.0001 to 1 mole, and preferably approximately
0.01 to 0.5 moles, based on the amount of the compound 4 (1 mole). The ligand is used
in an amount of generally approximately 0.0001 to 4 moles, and preferably approximately
0.01 to 2 moles, based on the amount of the compound represented by the formula 4
(1 mole).
[0144] Examples of the base may include organic amines (e.g., trimethylamine, triethylamine,
diisopropylethylamine, N-methylmorpholine, 1,8-diazabicyclo[5,4,0]undec-7-ene, pyridine,
N,N-dimethylaniline, etc.), alkaline metal salts (e.g., sodium hydrogen carbonate,
potassium hydrogen carbonate, sodium carbonate, potassium carbonate, cesium carbonate,
sodium phosphate, potassium phosphate, sodium hydroxide, potassium hydroxide, etc.),
metal hydrides (e.g., potassium hydride, sodium hydride, etc.), alkaline metal alkoxides
(e.g., sodium methoxide, sodium ethoxide, sodium-tert-butoxide, potassium-tert-butoxide,
etc.), and alkaline metal disilazides (e.g., lithium disilazide, sodium disilazide,
potassium disilazide, etc.). Among others, alkaline metal salts such as potassium
carbonate, cesium carbonate, sodium phosphate, and potassium phosphate, alkaline metal
alkoxides such as sodium-tert-butoxide and potassium-tert-butoxide, organic amines
such as triethylamine and diisopropylethylamine, and the like are preferable. The
base is used in an amount of generally approximately 0.1 to 50 moles, and preferably
approximately 1 to 20 moles, based on the amount of the compound represented by the
formula 4 (1 mole).
[0145] The solvent is not particularly limited, as long as it does not affect the reaction.
Examples of the solvent may include hydrocarbons (e.g., benzene, toluene, xylene,
etc.), halogenated hydrocarbons (e.g., chloroform, 1,2-dichloroethane, etc.), nitriles
(e.g., acetonitrile, etc.), ethers (e.g., dimethoxyethane, tetrahydrofuran, etc.),
alcohols (e.g., methanol, ethanol, etc.), aprotic polar solvents (e.g., N,N-dimethylformamide,
dimethyl sulfoxide, hexamethylphosphoramide, N-methylpyrrolidone, etc.), water, and
mixtures thereof. The reaction time is 0.1 to 100 hours, and preferably 0.5 to 24
hours. Thereafter, the reaction temperature is 0°C to the temperature at which the
solvent is boiled, and preferably 0°C to 150° C.
[0146] After completion of this reaction, an ester form corresponding to the used alcohol,
or a mixture of the ester form and the compound represented by the formula 5 is subjected
to a hydrolysis reaction, so that it can be converted to the compound represented
by the formula 5.
[0147] As such a base, sodium hydrogen carbonate, sodium carbonate, potassium carbonate,
cesium carbonate, sodium hydroxide, potassium hydroxide, lithium hydroxide, or the
like is preferably used. The base is used in an amount of generally approximately
0.5 to 100 moles, and preferably approximately 1 to 10 moles, based on the amount
of the compound represented by the formula 4 (1 mole).
[0148] The solvent is not particularly limited, as long as it does not affect the reaction.
For example, water, methanol, ethanol, isopropanol, tetrahydrofuran, 1,4-dioxane,
N,N-dimethylformamide and the like can be used alone or in combination. The reaction
time is 0.1 to 100 hours, and preferably 0.5 to 24 hours. Thereafter, the reaction
temperature is 0°C to the temperature at which the solvent is boiled, and preferably
0°C to 100° C.
[0149] The thus obtained compound represented by the formula 5 can be isolated and purified
by known separation and purification means, or can be subjected to the subsequent
step without isolation and purification.
< Step 4 >
[0151] The "protective group for a carboxyl group" is not particularly limited, as long
as it has a protective function. Examples of the protective group for a carboxyl group
may include: lower alkyl groups, such as, for example, a methyl group, an ethyl group,
a propyl group, an isopropyl group, and a tert-butyl group; halo lower alkyl groups,
such as, for example, a 2,2,2-trichloroethyl group; lower alkenyl groups, such as,
for example, an allyl group; for example, a trimethylsilylethoxymethyl group; and
aralkyl groups, such as, for example, a benzyl group, a p-methoxybenzyl group, a p-nitrobenzyl
group, a benzhydryl group, and a trityl group. In particular, a methyl group, an ethyl
group, a tert-butyl group, an allyl group, a benzyl group, a p-methoxybenzyl group,
or a trimethylsilylethoxymethyl group is preferable.
[0152] In the present reaction, a protective group such as, for example, a tert-butyl ester
group, a methyl ester group, or an ethyl ester group, is preferably introduced.
[0153] The protective group agent used in the present reaction may be, for example, 2-tert-butyl-1,3-diisopropylisourea.
Such a protective group agent is used in an amount of generally approximately 1 to
50 moles, and preferably approximately 1 to 10 moles, based on the amount of the compound
represented by the formula 5 (1 mole).
[0154] The solvent is not particularly limited, as long as it does not affect the reaction.
Examples of the solvent may include hydrocarbons (e.g., benzene, toluene, xylene,
etc.), halogenated hydrocarbons (e.g., chloroform, 1,2-dichloroethane, etc.), nitriles
(e.g., acetonitrile, etc.), ethers (e.g., dimethoxyethane, tetrahydrofuran, tert-butyl
methyl ether, etc.), alcohols (e.g., methanol, ethanol, etc.), aprotic polar solvents
(e.g., N,N-dimethylformamide, dimethyl sulfoxide, hexamethylphosphoramide, etc.),
water, and mixtures thereof. The reaction time is 0.1 to 100 hours, and preferably
0.5 to 24 hours. Thereafter, the reaction temperature is 0°C to the temperature at
which the solvent is boiled, and preferably 0°C to 100° C.
[0155] The thus obtained compound represented by the formula 6 can be isolated and purified
by known separation and purification means, or can be subjected to the subsequent
step without isolation and purification.
< Step 5 >
[0156] This step is a method of obtaining a compound represented by the formula 7 (wherein
L
3 represents a halogen atom) by halogenating the compound represented by the formula
6. Halogenation can be carried out by a method using fluorine, chlorine, bromine,
iodine, etc., or by a method using N-chlorosuccinimide, N-bromosuccinimide, N-iodosuccinimide,
etc. In the present reaction, the method using N-chlorosuccinimide, N-bromosuccinimide,
N-iodosuccinimide, etc. is preferable.
[0157] N-chlorosuccinimide, N-bromosuccinimide, N-iodosuccinimide, etc. can be used in an
amount of 1 to 10 equivalents, and preferably 1 to 3 equivalents, based on the amount
of the compound represented by the formula 6 (1 mole).
[0158] The solvent is not particularly limited, as long as it does not affect the reaction.
Examples of the solvent may include hydrocarbons (e.g., benzene, toluene, xylene,
etc.), halogenated hydrocarbons (e.g., chloroform, 1,2-dichloroethane, etc.), nitriles
(e.g., acetonitrile, etc.), ethers (e.g., dimethoxyethane, tetrahydrofuran, etc.),
alcohols (e.g., methanol, ethanol, etc.), aprotic polar solvents (e.g., N,N-dimethylformamide,
dimethyl sulfoxide, hexamethylphosphoramide, etc.), water, and mixtures thereof. The
reaction time is 0.1 to 100 hours, and preferably 0.5 to 24 hours. Thereafter, the
reaction temperature is 0°C to the temperature at which the solvent is boiled, and
preferably 0°C to 100°.
[0159] The thus obtained compound represented by the formula 7 can be isolated and purified
by known separation and purification means, or can be subjected to the subsequent
step without isolation and purification.
< Step 6 >
[0161] The protective group may be, for example, tert-butyloxycarbonyl. When such a tert-butyloxycarbonyl
group is used, for example, as a protective group, deprotection is preferably carried
out under acidic conditions. Examples of the acid used herein may include hydrochloric
acid, acetic acid, trifluoroacetic acid, sulfuric acid, and tosylic acid.
[0162] The acid is preferably used in an amount of approximately 1 to 100 equivalents based
on the amount of the compound represented by the formula 7 (1 mole).
[0163] The solvent used in the reaction is not particularly limited, as long as it does
not affect the reaction. Examples of the solvent used herein may include alcohols
(e.g., methanol, etc.), hydrocarbons (e.g., benzene, toluene, xylene, etc.), halogenated
hydrocarbons (e.g., methylene chloride, chloroform, 1,2-dichloroethane, etc.), nitriles
(e.g., acetonitrile, etc.), ethers (e.g., dimethoxyethane, tetrahydrofuran, etc.),
aprotic polar solvents (e.g., N,N-dimethylformamide, dimethyl sulfoxide, hexamethylphosphoramide,
etc.), and mixtures thereof. The reaction time is 0.1 to 100 hours, and preferably
0.5 to 24 hours. Thereafter, the reaction temperature is 0°C to 100°C, and preferably
0°C to 50°.
[0164] The thus obtained compound represented by the formula 8 can be isolated and purified
by known separation and purification means, or can be subjected to the subsequent
step without isolation and purification.
< Step 7 >
[0165] This step is a method of obtaining a compound represented by the formula 9 by performing
an amidation reaction between an amino group of the compound represented by the formula
8 and acrylic acid halide or acrylic acid anhydride.
[0166] In the case of using acrylic acid halide or acrylic acid anhydride, such acrylic
acid halide or acrylic acid anhydride is used in an amount of generally approximately
0.5 to 10 moles, and preferably approximately 1 to 5 moles, based on the amount of
the compound represented by the formula 8 (1 mole). It is to be noted that the present
acrylic acid halide or acrylic acid anhydride can be obtained as a commercially available
product or can be produced according to a known method.
[0167] In addition, a base can be added, as necessary. Examples of the base may include
organic amines (e.g., trimethylamine, triethylamine, isopropylethylamine, diisopropylethylamine,
N-methylmorpholine, 1,8-diazabicyclo[5,4,0]undec-7-ene, pyridine, N,N-dimethylaniline,
etc.), alkaline metal salts (e.g., sodium hydrogen carbonate, potassium hydrogen carbonate,
sodium carbonate, potassium carbonate, cesium carbonate, sodium phosphate, potassium
phosphate, sodium hydroxide, potassium hydroxide, etc.), metal hydrides (e.g., potassium
hydride, sodium hydride, etc.), and alkaline metal alkoxides (e.g., sodium methoxide,
sodium ethoxide, sodium-tert-butoxide, potassium-tert-butoxide, etc.). The base is
used in an amount of generally approximately 1 to 100 moles, and preferably approximately
1 to 10 moles, based on the amount of the compound represented by the formula 8 (1
mole).
[0168] The solvent used in the reaction is not particularly limited, as long as it does
not affect the reaction. Examples of the solvent used herein may include alcohols
(e.g., methanol, etc.), hydrocarbons (e.g., benzene, toluene, xylene, etc.), halogenated
hydrocarbons (e.g., methylene chloride, chloroform, 1,2-dichloroethane, etc.), nitriles
(e.g., acetonitrile, etc.), ethers (e.g., dimethoxyethane, tetrahydrofuran, etc.),
aprotic polar solvents (e.g., N,N-dimethylformamide, dimethyl sulfoxide, hexamethylphosphoramide,
, etc.), and mixtures thereof. The reaction time is 0.1 to 100 hours, and preferably
0.5 to 24 hours. Thereafter, the reaction temperature is 0°C to the temperature at
which the solvent is boiled, and preferably 0°C to 100° C.
[0169] The thus obtained compound represented by the formula 9 can be isolated and purified
by known separation and purification means, or can be subjected to the subsequent
step without isolation and purification.
< Step 8 >
[0170] This step is a method of obtaining a compound represented by the formula 10 by performing
a Sonogashira reaction between the compound represented by the formula 9 and an acetylene
derivative that is a commercially available product or can be produced by a known
method.
[0171] The acetylene derivative can be used in an amount of 1 to 50 equivalents, and preferably
1 to 10 equivalents, based on the amount of the compound represented by the formula
9 (1 mole).
[0172] Examples of the transition metal catalyst used herein may include palladium catalysts
(e.g., palladium acetate, palladium chloride, tetrakistriphenylphosphinepalladium,
dichlorobis(triphenylphosphine)palladium, dichlorobis(triphenylphosphine)dipalladium,
etc.), and nickel catalysts (e.g., nickel chloride, etc.). As necessary, a ligand
(e.g., triphenylphosphine, tri-tert-butylphosphine, etc.) may be added, and a copper
catalyst (e.g., copper iodide, copper bromide, or copper chloride) or the like may
be used as a co-catalyst. The amount of the transition metal catalyst used is different
depending on the type of the catalyst. The transition metal catalyst is used in an
amount of generally approximately 0.0001 to 1 mole, and preferably approximately 0.01
to 0.5 moles, based on the amount of the compound represented by the formula 9 (1
mole). The ligand is used in an amount of generally approximately 0.0001 to 4 moles,
and preferably approximately 0.01 to 2 moles, based on the amount of the compound
represented by the formula 9 (1 mole). The copper catalyst is used in an amount of
generally approximately 0.0001 to 4 moles, and preferably approximately 0.010 to 2
moles, based on the amount of the compound represented by the formula 9 (1 mole).
[0173] Examples of the base may include organic amines (e.g., trimethylamine, triethylamine,
diisopropylethylamine, N-methylmorpholine, 1,8-diazabicyclo[5,4,0]undec-7-ene, pyridine,
N,N-dimethylaniline, etc.), alkaline metal salts (e.g., sodium hydrogen carbonate,
potassium hydrogen carbonate, sodium carbonate, potassium carbonate, cesium carbonate,
sodium phosphate, potassium phosphate, sodium hydroxide, potassium hydroxide, etc.),
metal hydrides (e.g., potassium hydride, sodium hydride, etc.), alkaline metal alkoxides
(e.g., sodium methoxide, sodium ethoxide, sodium-tert-butoxide, potassium-tert-butoxide,
etc.), and alkaline metal disilazide (e.g., lithium disilazide, sodium disilazide,
potassium disilazide, etc.). Among these, preferred examples of the base may include:
alkaline metal salts, such as potassium carbonate, cesium carbonate, sodium phosphate,
and potassium phosphate; alkaline metal alkoxides, such as sodium-tert-butoxide and
potassium-tert-butoxide; and organic amines, such as triethylamine and diisopropylethylamine.
The base is used in an amount of generally approximately 0.1 to 10 moles, and preferably
approximately 1 to 5 moles, based on the amount of the compound represented by the
formula 9 (1 mole).
[0174] The solvent is not particularly limited, as long as it does not affect the reaction.
Examples of the solvent may include hydrocarbons (e.g., benzene, toluene, xylene,
etc.), halogenated hydrocarbons (e.g., chloroform, 1,2-dichloroethane, etc.), nitriles
(e.g., acetonitrile, etc.), ethers (e.g., dimethoxyethane, tetrahydrofuran, etc.),
alcohols (e.g., methanol, ethanol, etc.), aprotic polar solvents (e.g., N,N-dimethylformamide,
dimethyl sulfoxide, hexamethylphosphoramide, etc.), water, and mixtures thereof. The
reaction time is 0.1 to 100 hours, and preferably 0.5 to 24 hours. Thereafter, the
reaction temperature is 0°C to the temperature at which the solvent is boiled, and
preferably 0°C to 150° C.
[0175] The thus obtained compound represented by the formula 10 can be isolated and purified
by known separation and purification means, or can be subjected to the subsequent
step without isolation and purification.
< Step 9 >
[0177] The protective group may be, for example, tert-butyl ester. When such a tert-butyl
ester group is used as a protective group, for example, deprotection is preferably
carried out under acidic conditions. Examples of the acid used herein may include
hydrochloric acid, acetic acid, trifluoroacetic acid, sulfuric acid, and tosylic acid.
[0178] The acid is preferably used in an amount of approximately 1 to 100 equivalents based
on the amount of the compound represented by the formula 10 (1 mole).
[0179] The solvent used in the reaction is not particularly limited, as long as it does
not affect the reaction. Examples of the solvent used herein may include alcohols
(e.g., methanol, etc.), hydrocarbons (e.g., benzene, toluene, xylene, etc.), halogenated
hydrocarbons (e.g., methylene chloride, chloroform, 1,2-dichloroethane, etc.), nitriles
(e.g., acetonitrile, etc.), ethers (e.g., dimethoxyethane, tetrahydrofuran, etc.),
aprotic polar solvents (e.g., N,N-dimethylformamide, dimethyl sulfoxide, hexamethylphosphoramide,
, etc.), and mixtures thereof. The reaction time is 0.1 to 100 hours, and preferably
0.5 to 24 hours. Thereafter, the reaction temperature is 0°C to 100°C, and preferably
0°C to 50°.
[0180] The thus obtained compound represented by the formula 11 can be isolated and purified
by known separation and purification means, or can be subjected to the subsequent
step without isolation and purification.
< Step 10 >
[0181] This step is a method of obtaining a compound represented by the formula (I) by performing
an amidation reaction between a carboxyl group of the compound represented by the
formula 11 and an amine that is a commercially available product or can be produced
by a known method.
[0183] Examples of the condensing agent may include N,N'-dicyclohexylcarbodiimide (DCC),
N,N'-diisopropylcarbodiimide (DIC), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide
hydrochloride (WSC), diphenylphosphoryl azide (DPPA), benzotriazol-1-yl-oxytrisdimethylaminophosphonium
hexafluorophosphate (BOP), benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate
(PyBOP), 7-azabenzotriazol-1-yloxytrispyrrolidinophosphonium phosphate (PyAOP), bromotrispyrrolidinophosphonium
hexafluorophosphate (BroP), chlorotris(pyrrolidin-1-yl)phosphonium hexafluorophosphate
(PyCroP), 3-(diethoxyphosphoryloxy)-1,2,3-benzotriazin-4(3H)-one (DEPBT), O-(azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate (HATU), and 4-(5,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholine
hydrochloride (DMTMM). Examples of the additive used at that time may include 1-hydroxybenzotriazole
(HOBt), 1-hydroxy-7-azabenzotriazole (HOAt), and N-hydroxysuccinimide (HOSu). Such
agents are used in an amount of generally approximately 1 to 100 moles, and preferably
approximately 1 to 10 moles, based on the amount of the compound represented by the
formula 11 (1 mole).
[0184] In addition, as necessary, a base can be added. Examples of such a base may include
organic amines (e.g., trimethylamine, triethylamine, diisopropylethylamine, N-methylmorpholine,
1,8-diazabicyclo[5,4,0]undec-7-ene, pyridine, N,N-dimethylaniline, etc.), alkaline
metal salts (e.g., sodium hydrogen carbonate, potassium hydrogen carbonate, sodium
carbonate, potassium carbonate, cesium carbonate, sodium phosphate, potassium phosphate,
sodium hydroxide, potassium hydroxide, etc.), metal hydrides (e.g., potassium hydride,
sodium hydride, etc.), and alkaline metal alkoxides (e.g., sodium methoxide, sodium
ethoxide, sodium-tert-butoxide, potassium-tert-butoxide, etc.). The base is used in
an amount of generally approximately 1 to 100 moles, and preferably approximately
1 to 10 moles, based on the amount of the compound represented by the formula 11 (1
mole).
[0185] The solvent used in the reaction is not particularly limited, as long as it does
not affect the reaction. Examples of the solvent used herein may include alcohols
(e.g., methanol, etc.), hydrocarbons (e.g., benzene, toluene, xylene, etc.), halogenated
hydrocarbons (e.g., methylene chloride, chloroform, 1,2-dichloroethane, etc.), nitriles
(e.g., acetonitrile, etc.), ethers (e.g., dimethoxyethane, tetrahydrofuran, etc.),
aprotic polar solvents (e.g., N,N-dimethylformamide, dimethyl sulfoxide, hexamethylphosphoramide,
, etc.), and mixtures thereof. The reaction time is 0.1 to 100 hours, and preferably
0.5 to 24 hours. Thereafter, the reaction temperature is 0°C to the temperature at
which the solvent is boiled, and preferably 0°C to 100° C.
[0186] The thus obtained compounds (I) and (II) can be isolated and purified according to
known separation and purification means, such as, for example, concentration , vacuum
concentration, crystallization, solvent extraction, reprecipitation, or chromatography.
[0187] In the above-described production method, the steps ranging from the "introduction
of a protective group into a carboxyl group of the compound represented by the formula
5" (Step 4) to the "amidation reaction between a carboxyl group of the compound represented
by the formula 11 and an amine that is a commercially available product or can be
produced by a known method" (Step 10) are successively carried out in this order.
However, the order of performing these steps can be changed. Moreover, the "introduction
of a protective group into a carboxyl group of the compound represented by the formula
5" (Step 4) and the "removal of the protective group for a carboxy group from the
compound represented by the formula 10" (Step 9) can be omitted.
[0188] Specifically, individual steps are carried out in the order of the "amidation reaction
between a carboxyl group of the compound represented by the formula 11 and an amine
that is a commercially available product or can be produced by a known method" (Step
10), the "halogenation of the compound represented by the formula 6" (Step 5), the
"removal of the protective group for an amino group from the compound represented
by the formula 7" (Step 6), the "amidation reaction between an amino group of the
compound represented by the formula 8 and acrylic acid halide or acrylic acid anhydride"
(Step 7), and the "Sonogashira reaction between the compound represented by the formula
9 and an acetylene derivative that is a commercially available product or can be produced
by a known method, when L3 of the compound represented by the formula 9 has a leaving
group such as halogen" (Step 8), so that the concerned compound can be induced to
the compounds represented by the formulae (I) and (II). The conditions applied in
individual steps are the same as those as described above.
[0189] When the compound of the present invention has an isomer, such as an optical isomer,
a stereoisomer, a rotational isomer, or a tautomer, all of these isomers or mixtures
thereof are included in the compound of the present invention, unless otherwise stated.
For example, when the compound of the present invention has an optical isomer, both
a racemate, and an optical isomer obtained as a result of racemic resolution are included
in the compound of the present invention, unless otherwise stated.
[0190] The salt of the compound of the present invention means a pharmaceutically acceptable
salt, and it may be, for example, a base-added salt or an acid-added salt.
[0191] The pyrimidine compound of the present invention or a salt thereof also includes
a prodrug. The "prodrug" means a compound that is converted to the compound of the
present invention or a salt thereof as a result of the reaction with an enzyme, stomach
acid, etc. under physiological conditions in a living body; namely, a compound that
enzymatically causes oxidation, reduction, hydrolysis, etc., so that it is converted
to the compound of the present invention or a salt thereof, or a compound that causes
hydrolysis, etc. with stomach acid or the like, so that it is converted to the compound
of the present invention or a salt thereof. Otherwise, it may also be a compound that
is converted to the compound of the present invention or a salt thereof under physiological
conditions as described in "
Iyakuhin no Kaihatsu (Development of Pharmaceutical Products)," Hirokawa Shoten, 1990,
Vol. 7, Bunshi Sekkei (Molecular Designing), pp. 163 to 198.
[0192] The pyrimidine compound of the present invention or a salt thereof may be an amorphous
material or a crystal. Although the crystal form thereof may be a single crystal or
a polymorphic mixture, they are included in the compound of the present invention
or a salt thereof. The crystal can be produced by crystallizing the compound of the
present invention or a salt thereof, applying a known crystallization method. The
compound of the present invention or a salt thereof may be either a solvate (e.g.,
a hydrate, etc.), or a non-solvate, and both of them are included in the compound
of the present invention or a salt thereof. Compounds labeled with radioisotopes (e.g.,
3H,
14C,
35S,
125I, etc.) and the like are also included in the compound of the present invention or
a salt thereof.
[0193] The compound of the present invention or a salt thereof has excellent EGFR inhibitory
activity. Accordingly, the compound of the present invention or a salt thereof is
useful as an antitumor agent against malignant tumor having EGFR overexpression, EGFR
gene amplification, or EGFR mutation, etc. In addition, since significant weight reduction
was not found in mice, the present compound or a salt thereof is advantageous in that
it has a few side effects.
[0194] In the present description, the term "EGFR" includes the EGFR of a human or a non-human
mammal, and it is preferably human EGFR. The NCBI Gene ID for the human EGFR is 1956.
Furthermore, the term "EGFR" includes isoforms.
[0195] Since the compound of the present invention or a salt thereof has excellent EGFR
inhibitory activity, it is useful as a medicament for preventing or treating disease
associated with EGFR.
[0196] The "disease associated with EGFR" means disease, in which a reduction in the incidence,
or the remission, alleviation and/or complete recovery of the symptoms thereof is
achieved by deleting, suppressing and/or inhibiting the function of EGFR. Examples
of such disease may include malignant tumors, but are not limited thereto. Preferred
examples of the disease may include malignant tumors having EGFR overexpression, EGFR
gene amplification, or EGFR mutation.
[0197] One embodiment of the present invention provides a therapeutic agent for a disease
associated with EGFR, comprising the compound of the present invention or a salt thereof.
One embodiment of the present invention provides an EGFR inhibitor comprising the
compound of the present invention or a salt thereof. One embodiment of the present
invention provides a therapeutic agent for EGFR-positive tumor, comprising the compound
of the present invention or a salt thereof. One embodiment of the present invention
provides the compound of the present invention or a salt thereof for treating a disease
associated with EGFR. One embodiment of the present invention provides use of the
compound of the present invention or a salt thereof for treating a disease associated
with EGFR. One embodiment of the present invention provides use of the compound of
the present invention or a salt thereof for producing a therapeutic agent for a disease
associated with EGFR. One embodiment of the present invention provides a method for
treating a disease associated with EGFR, comprising administering an effective amount
of the compound of the present invention or a salt thereof to a subject in need thereof.
One embodiment of the present invention provides the compound of the present invention
or a salt thereof for treating EGFR-positive tumor. One embodiment of the present
invention provides use of the compound of the present invention or a salt thereof
for treating EGFR-positive tumor. One embodiment of the present invention provides
use of the compound of the present invention or a salt thereof for producing a therapeutic
agent for EGFR-positive tumor. One embodiment of the present invention provides a
method for treating EGFR-positive tumor, comprising administering an effective amount
of the compound of the present invention or a salt thereof to a subject in need thereof.
[0198] The compound according to one embodiment of the present invention or a salt thereof
inhibits wild-type EGFR, and mutant EGFR having an insertion mutation, a point mutation,
or a deletion mutation, etc. One embodiment of the present invention provides a compound
having inhibitory activity against wild-type EGFR and mutant EGFR, or a salt thereof,
or a medicament or a pharmaceutical composition comprising the same. One embodiment
of the present invention provides an inhibitor for wild-type EGFR and mutant EGFR,
comprising the compound of the present invention or a salt thereof. One embodiment
of the present invention provides a method for inhibiting wild-type EGFR and mutant
EGFR, comprising administering an effective amount of the compound of the present
invention or a salt thereof to a subject in need thereof. One embodiment of the present
invention provides use of the compound of the present invention or a salt thereof
for producing an inhibitor for wild-type EGFR and mutant EGFR. One embodiment of the
present invention provides the compound of the present invention or a salt thereof
for use as an inhibitor for wild-type EGFR and mutant EGFR. One embodiment of the
present invention provides use of the compound of the present invention or a salt
thereof for inhibiting wild-type EGFR and mutant EGFR. Another embodiment of the present
invention provides use of the compound of the present invention or a salt thereof
for preventing or treating diseases associated with wild-type EGFR and mutant EGFR.
[0199] The human wild-type EGFR gene is shown in, for example, SEQ ID NO: 1. The human wild-type
EGFR protein consists of the amino acid sequence set forth in, for example, SEQ ID
NO: 2. The nucleotide sequence information of the human wild-type EGFR gene and the
amino acid sequence information of the human wild-type EGFR protein can be obtained
from, for example, NCBI Reference Sequence: NM_005228 and NCBI Reference Sequence:
NP_005219, respectively.
[0200] In several embodiments, the pyrimidine compound of the present invention or a salt
thereof exhibits inhibitory activity against mutant EGFR. In the present description,
"mutant EGFR" is EGFR having one or more activating mutations or resistance acquiring
mutations, such as insertion mutations point mutations, or deletion mutations, in
exon 18 region, exon 19 region, exon 20 region, exon 21 region or the like of human
wild-type EGFR.
[0201] In the present description, "exon 18" corresponds to a region from positions 688
to 728 in the amino acid sequence of the human wild-type EGFR protein (e.g., a protein
consisting of the amino acid sequence set forth in SEQ ID NO: 2).
[0202] In the present invention, "exon 18 mutation" refers to a point mutation or a deletion
mutation in exon 18 region resulting in an amino acid mutation in the human wild-type
EGFR protein (e.g., a protein consisting of the amino acid sequence set forth in SEQ
ID NO: 2). Examples of the point mutation in exon 18 include point mutation E709X
or G719X in exon 18 region, which substitutes glutamic acid at position 709 or glycine
at position 719 with any amino acid. Examples of E709X include point mutation E709K
in exon 18 region, which substitutes glutamic acid at position 709 with lysine, and
point mutation E709A in exon 18 region, which substitutes glutamic acid at position
709 with alanine. Examples of G719X include point mutation G719A in exon 18 region,
which substitutes glycine at position 719 with alanine, point mutation G719S in exon
18 region, which substitutes glycine at position 719 with serine, and point mutation
G719C in exon 18 region, which substitutes glycine at position 719 with cysteine.
The deletion mutation in exon 18 region encompasses not only a mutation in exon 18
region, which deletes some amino acids, but also a mutation therein, which inserts
any one or more amino acids in addition to the amino acid deletion. Examples of the
deletion mutation in exon 18 include a mutation in exon 18 region, which deletes glutamic
acid at position 709 and threonine at position 710 and then inserts aspartic acid
(Del E709-T710insD).
[0203] In the present description, "exon 19" corresponds to a region from positions 729
to 761 in the amino acid sequence of the human wild-type EGFR protein (e.g., a protein
consisting of the amino acid sequence set forth in SEQ ID NO: 2).
[0204] In the present description, "exon 19 mutation" refers to a mutation in exon 19 region,
which deletes one or more amino acids in the human wild-type EGFR protein (e.g., a
protein consisting of the amino acid sequence set forth in SEQ ID NO: 2). The deletion
mutation in exon 19 region encompasses not only a mutation in exon 19 region, which
deletes some amino acids, but also a mutation therein, which inserts any one or more
amino acids in addition to the amino acid deletion. Examples of the exon 19 deletion
mutation include a mutation in exon 19 region, which deletes 5 amino acids from glutamic
acid at position 746 to alanine at position 750 (Del E746-A750 (or also referred to
as d746-750)), a mutation in exon 19 region, which deletes 7 amino acids from leucine
at position 747 to proline at position 753 and then inserts serine (Del L747-P753insS),
a mutation in exon 19 region, which deletes 5 amino acids from leucine at position
747 to threonine at position 751 (Del L747-T751), and a mutation in exon 19 region,
which deletes 4 amino acids from leucine at position 747 to alanine at position 750
and then inserts proline (Del L747-A750insP). In one preferred embodiment of the present
invention, the exon 19 deletion mutation is a mutation in exon 19 region, which deletes
5 amino acids from glutamic acid at position 746 to alanine at position 750 (Del E746-A750).
[0205] In the present description, "exon 20" corresponds to a region from positions 762
to 823 in the amino acid sequence of the human wild-type EGFR protein (e.g., a protein
consisting of the amino acid sequence set forth in SEQ ID NO: 2).
[0206] In the present invention, "exon 20 mutation" refers to a point mutation, an insertion
mutation, a deletion mutation, or the like in exon 20 region resulting in an amino
acid mutation in the human wild-type EGFR protein (e.g., a protein consisting of the
amino acid sequence set forth in SEQ ID NO: 2). Examples of the exon 20 mutation include
A763insFQEA, A767insASV, S768dupSVD, V769insASV, D770insNPG, D770insSVD, and D773insNPH
(Nature medicine,24,p638-646,2018). In one preferred embodiment of the present invention,
the exon 20 mutation is one or more insertion mutations or point mutations selected
from V769_D770insASV, D770_N771insNPG, D770_N771insSVD, H773_V774insNPH, and T790M.
[0207] In the present description, "exon 21" corresponds to a region from positions 824
to 875 in the amino acid sequence of the human wild-type EGFR protein (e.g., a protein
consisting of the amino acid sequence set forth in SEQ ID NO: 2).
[0208] In the present invention, "exon 21 mutation" refers to a point mutation in exon 21
region resulting in an amino acid mutation in the human wild-type EGFR protein (e.g.,
a protein consisting of the amino acid sequence set forth in SEQ ID NO: 2). Examples
of the point mutation in exon 21 include point mutations in exon 21 region, which
substitute one amino acid and preferably include position mutation L858X or L861X
in exon 21 region, which substitutes leucine at position 858 or leucine at position
861 with any amino acid. Examples of L858X include point mutation L858R in exon 21
region, which substitutes leucine at position 858 with arginine. Examples of L861X
include point mutation L861Q in exon 21 region, which substitutes leucine at position
861 with glutamine. In one preferred embodiment of the present invention, the point
mutation in exon 21 is L858R.
[0209] In several embodiments, with regard to a mutation in a certain EGFR isoform, even
when position of the mutation is different from position of an amino acid shown in
SEQ ID NO: 2 due to deletion or insertion of an amino acid(s), it is understood that
the mutation is the same as the mutation at a position corresponding to position of
the amino acid shown in SEQ ID NO: 2. Hence, for example, the threonine at position
790 in the EGFR shown in SEQ ID NO: 2 corresponds to threonine at position 745 in
EGFR consisting of the amino acid sequence set forth in SEQ ID NO: 4. For example,
the term "T790M" means that the threonine at position 790 in the EGFR shown in SEQ
ID NO: 2 is mutated to methionine. Since such "T790M" is at a position corresponding
to the amino acid at position 745 in EGFR consisting of the amino acid sequence set
forth in SEQ ID NO: 4, "T745M" in the EGFR consisting of the amino acid sequence set
forth in SEQ ID NO: 4 corresponds to "T790M" in the EGFR shown in SEQ ID NO: 2. For
example, the threonine at position 790 in the EGFR shown in SEQ ID NO: 2 corresponds
to threonine at position 523 in EGFR consisting of the amino acid sequence set forth
in SEQ ID NO: 6. For example, the term "T790M" means that the threonine at position
790 in the EGFR shown in SEQ ID NO: 2 is mutated to methionine. Since such "T790M"
is at a position corresponding to the amino acid at position 523 in EGFR consisting
of the amino acid sequence set forth in SEQ ID NO: 6, "T523M" in the EGFR consisting
of the amino acid sequence set forth in SEQ ID NO: 6 corresponds to "T790M" in the
EGFR shown in SEQ ID NO: 2. Besides, the position of an amino acid in SEQ ID NO: 2
that corresponds to a certain amino acid in a certain EGFR isoform can be confirmed
by Multiple Alignment of BLAST.
[0210] SEQ ID NOs: 1 to 6 are as described below.
EGFR variant 1
EGFR variant 5
EGFRvIII (del2-7 EGFR)
[0214] In the present invention, "EGFR-positive tumor" is tumor in which the EGFR protein
or the EGFR gene is detected. The EGFR protein and the EGFR gene also include mutant
EGFR protein and mutant EGFR gene having a point mutation, an insertion mutation,
or a deletion mutation, etc.
[0215] Examples of the method for detecting the EGFR protein include usual detection methods
commonly used, such as ELISA, Western blotting, and immunostaining using an antibody
specifically binding to the EGFR protein. The antibody specifically binding to the
EGFR protein may be a commercially available product or may be prepared by a usual
method commonly used.
[0216] Examples of the method for detecting the EGFR gene include usual detection methods
commonly used, such as Northern blotting, Southern blotting, RT-PCR, real-time PCR,
digital PCR, DNA microarrays,
in situ hybridization, and sequence analysis. Another example thereof includes a detection
method using cobas EGFR mutation detection kit (Roche Diagnostics K.K.), which is
a commercially available EGFR gene mutation detection kit.
[0217] In the present description, the term "effective amount" used regarding the pyrimidine
compound of the present invention means the amount of the compound of the present
invention that induces the biological or medical response of a subject, such as, for
example, reduction or inhibition of enzyme or protein activity; or ameliorates symptoms,
alleviates conditions, and retards or delays the progression of disease; or prevents
disease; etc. (therapeutically effective amount).
[0218] In the present description, the term "subject" includes mammals and non-mammals.
Examples of the mammal may include, but are not limited to, a human, a chimpanzee,
an ape, a monkey, a bovine, a horse, sheep, a goat, a swine, a rabbit, a dog, a cat,
a rat, a mouse, a Guinea pig, a hedgehog, a kangaroo, a mole, a wild pig, a bear,
a tiger, and a lion. Examples of the non-mammal may include, but are not limited to,
birds, fish, and reptiles. In one embodiment, the subject is a human, and may be a
human who has been diagnosed to need the treatment for the symptoms, conditions or
disease disclosed in the present description.
[0219] Upon the use of the compound of the present invention or a salt thereof as a medicament,
a pharmaceutically acceptable carrier is mixed into it, as necessary, and various
types of dosage forms can be adopted depending on the preventive or therapeutic purpose.
Examples of the dosage form may include all of an oral agent, an injection, a suppository,
an ointment, and a patch. Preferably, an oral agent is adopted. These dosage forms
can be produced by commonly used production methods that are known to skilled persons.
[0220] One embodiment of the present invention provides an antitumor agent for oral administration,
comprising the compound of the present invention or a salt thereof as an active ingredient.
In addition, one embodiment of the present invention provides a method for preventing
and/or treating tumor, comprising administering an effective amount of the compound
of the present invention or a salt thereof to a subject in need thereof by oral administration.
Moreover, one embodiment of the present invention provides use of the compound of
the present invention or a salt thereof for the production of an antitumor agent for
oral administration. Furthermore, one embodiment of the present invention provides
the compound of the present invention or a salt thereof for use in the prevention
and/or treatment of tumor by oral administration thereof.
[0221] One embodiment of the present invention provides a pharmaceutical composition comprising
the compound of the present invention or a salt thereof. The pharmaceutical composition
according to one embodiment of the present invention comprises the compound of the
present invention or a salt thereof, and a pharmaceutically acceptable carrier. Further,
one embodiment of the present invention provides use of the compound of the present
invention or a salt thereof for the production of a pharmaceutical composition. Another
embodiment of the present invention provides the compound of the present invention
or a salt thereof for use as a medicament.
[0222] As pharmaceutically acceptable carriers, various types of organic or inorganic carrier
substances, which are commonly used as preparation materials, are used. When the compound
of the present invention is processed into a solid preparation, examples of the pharmaceutically
acceptable carrier mixed into the compound of the present invention may include an
excipient, a binder, a disintegrator, a lubricant, a coating agent, and a coloring
agent. When the compound of the present invention is processed into a liquid preparation,
examples of the pharmaceutically acceptable carrier mixed into the compound of the
present invention may include a solvent, a solubilizer, a suspending agent, a tonicity
agent, a buffer, and a soothing agent. In addition, preparation additives such as
an antiseptic, an antioxidant, a sweetener, and a stabilizer can also be used, as
necessary.
[0223] In the case of preparing a solid preparation for oral administration, an excipient,
and as necessary, a binder, a disintegrator, a lubricant, a coloring agent, a corrigent,
etc. are added to the pyrimidine compound of the present invention, and thereafter,
a tablet, a coated tablet, a granule, a powder agent, a capsule, etc. can be produced
according to ordinary methods.
[0224] In the case of preparing an injection, a pH adjuster, a buffer, a stabilizer, a tonicity
agent, a local anesthetic, etc. are added to the pyrimidine compound of the present
invention, and thereafter, subcutaneous, intramuscular, and intravenous injections
can be produced according to ordinary methods.
[0225] The amount of the compound of the present invention to be mixed into the above-described
each dosage unit form depends on the symptoms of a subject to whom the present pyrimidine
compound should be applied, the dosage form and the like, and thus, the amount of
the compound of the present invention is not constant. In general, it is preferable
that the applied dose is set to be 0.05 to 1000 mg per dosage unit form in the case
of an oral agent, it is set to be 0.01 to 500 mg per dosage unit form in the case
of an injection, and it is set to be 1 to 1000 mg per dosage unit form in the case
of a suppository.
[0226] The daily dose of a drug having the above-described dosage form is different depending
on the symptoms, body weight, age, sex and the like of a subject, and thus, it cannot
be generally determined. However, the compound of the present invention may be administered
to an adult (body weight: 50 kg) at a daily dose of generally 0.05 to 5000 mg, and
preferably 0.1 to 1000 mg.
[0227] The malignant tumor that is the target of the present invention is not particularly
limited. Examples of the tumor may include brain tumor, head and neck cancer, digestive
cancer (esophageal cancer, stomach cancer, duodenal cancer, liver cancer, biliary
tract cancer (gallbladder and/or bile duct cancer, etc.), pancreatic cancer, colorectal
cancer (colon cancer, rectal cancer, etc.), etc.), lung cancer (non-small cell lung
cancer, small cell lung cancer, mesothelioma, etc.), breast cancer, genital cancer
(ovarian cancer, uterine cancer (cervical cancer, endometrial cancer, etc.), etc.),
urinary organ cancer (kidney cancer, bladder cancer, prostate cancer, testicular tumor,
etc.), hematopoietic tumor (leukemia, malignant lymphoma, multiple myeloma, etc.),
bone and/or soft tissue tumor, and skin cancer. Among these, preferable is lung cancer,
breast cancer, stomach cancer, head and neck cancer, brain tumor, colorectal cancer,
bladder cancer, biliary tract cancer or uterine cancer, and more preferable is lung
cancer, breast cancer, colorectal cancer, or brain tumor.
[0228] In one embodiment, the tumor that is the target of the present invention is a malignant
tumor having EGFR overexpression, EGFR gene amplification, or EGFR mutation. Specific
examples of the tumor may include brain tumor, head and neck cancer, digestive cancer
(esophageal cancer, stomach cancer, duodenal cancer, liver cancer, biliary tract cancer
(gallbladder and/or bile duct cancer, etc.), pancreatic cancer, colorectal cancer
(colon cancer, rectal cancer, etc.), etc.), lung cancer (non-small cell lung cancer,
small cell lung cancer, mesothelioma, etc.), breast cancer, genital cancer (ovarian
cancer, uterine cancer (cervical cancer, endometrial cancer, etc.), etc.), urinary
organ cancer (kidney cancer, bladder cancer, prostate cancer, testicular tumor, etc.),
hematopoietic tumor (leukemia, malignant lymphoma, multiple myeloma, etc.), bone and/or
soft tissue tumor, and skin cancer. Among these, preferable is lung cancer, breast
cancer, stomach cancer, head and neck cancer, brain tumor, colorectal cancer, bladder
cancer, biliary tract cancer or uterine cancer, and more preferable is lung cancer,
breast cancer, colorectal cancer, or brain tumor.
[0229] In one embodiment, the tumor that is the target of the present invention is an EGFR
positive tumor. Specific examples of the tumor may include brain tumor, head and neck
cancer, digestive cancer (esophageal cancer, stomach cancer, duodenal cancer, liver
cancer, biliary tract cancer (gallbladder and/or bile duct cancer, etc.), pancreatic
cancer, colorectal cancer (colon cancer, rectal cancer, etc.), etc.), lung cancer
(non-small cell lung cancer, small cell lung cancer, mesothelioma, etc.), breast cancer,
genital cancer (ovarian cancer, uterine cancer (cervical cancer, endometrial cancer,
etc.), etc.), urinary organ cancer (kidney cancer, bladder cancer, prostate cancer,
testicular tumor, etc.), hematopoietic tumor (leukemia, malignant lymphoma, multiple
myeloma, etc.), bone and/or soft tissue tumor, and skin cancer. Among these, preferable
is lung cancer, breast cancer, stomach cancer, head and neck cancer, brain tumor,
colorectal cancer, bladder cancer, biliary tract cancer or uterine cancer, and more
preferable is lung cancer, breast cancer, colorectal cancer, or brain tumor.
[0230] In one embodiment, the tumor is a brain tumor. The pyrimidine compound of the present
invention may be useful for the treatment of the symptoms of brain that is required
to pass through the blood-brain barrier. The pyrimidine compound of one embodiment
has favorable permeability through the blood-brain barrier for the delivery thereof
into the brain, namely, excellent brain penetration properties. As an indicator of
the penetration properties of the compound into the brain, the concentration of the
compound in the brain or a Kp value (brain-to-plasma drug concentration ratio) is
applied.
[0231] The brain tumor treated with the pyrimidine compound of the present invention includes
metastatic brain tumor and primary brain tumor.
[0232] Examples of the brain tumor may include, but are not particularly limited to, metastatic
brain tumor (e.g., brain metastasis of lung cancer, breast cancer, stomach cancer,
colorectal cancer, bladder cancer, biliary tract cancer, uterine cancer, etc. (preferably,
lung cancer, breast cancer, or stomach cancer)), piliocytic astrocytoma, diffuse astrocytoma,
oligodendroma and/or oligodendroastrocytoma, anaplastic astrocytoma and/or anaplastic
oligodendroglioma, anaplastic oligodendroastrocytoma, glioblastoma, ependymoma, anaplastic
ependymoma, ganglioglioma, central neurocytoma, medulloblastoma, germinoma, central
nervous system malignant lymphoma, meningioma, neurilemmoma, GH secreting pituitary
adenoma, PRL-secreting pituitary adenoma, ACTH-secreting pituitary adenoma, nonfunctional
pituitary adenoma, craniopharyngioma, chordoma, hemangioblastoma, and epidermoid tumor.
EXAMPLES
[0233] Hereinafter, the present invention will be described in detail in the following examples.
However, these examples are not intended to limit the scope of the present invention.
[0234] In the present description, "room temperature" generally means a temperature that
is from approximately 10°C to approximately 35°C. In addition, in the following Examples
regarding compounds, "%" indicates weight percent, unless otherwise specified.
[0235] Various types of reagents used in the Examples were commercially available products,
unless otherwise specified. Silica gel chromatography was carried out using Biotage
SNAP Cartridge Ultra, manufactured by Biotage Japan Ltd. Basic silica gel chromatography
was carried out using Biotage SNAP Cartridge Isolute Flash-NH2, manufactured by Biotage
Japan Ltd.
[0236] Preparative thin-layer chromatography was carried out using Kieselgel TM60F254, Art.
5744, manufactured by Merck, or NH2 Silica Gel 60F254 Plate-Wako, manufactured by
FUJIFILM Wako Pure Chemical Industries, Ltd.
[0237] 1H-NMR was measured using tetramethylsilane as a reference material, and employing
AL400 (400 MHz) manufactured by JEOL, Mercury (400 MHz) manufactured by Varian, or
Inova (400 MHz) manufactured by Varian. Moreover, mass spectrum was measured using
Micromass ZQ or SQD manufactured by Waters, according to electrospray ionization (ESI)
or atmospheric pressure chemical ionization (APCI). Microwave reaction was carried
out using Initiator manufactured by Biotage Japan Ltd.
[0238] Abbreviations have the following meanings.
s: Singlet
d: Doublet
t: Triplet
q: Quartet
dd: Double doublet
dt: Double triplet
td: Triple doublet
tt: Triple triplet
ddd: Double double doublet
ddt: Double double triplet
dtd: Double triple doublet
tdd: Triple double doublet
m: Multiplet
br: Broad
ATP: Adenosine triphosphate
DMSO-d6: Deuterated dimethyl sulfoxide
CDCl3: Deuterated chloroform
EDTA: Ethylenediaminetetraacetic acid
THF: Tetrahydrofuran
DMF: N,N-dimethylformamide
DMSO: Dimethyl sulfoxide
NMP: N-methyl pyrrolidone
HATU: O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate
HPMC: Hypromellose
PdCl2(PPh3)2: Dichlorobis(triphenylphosphine)palladium(II)
< Reference Example 1 >
Reference Example 1(1)
tert-Butyl (2S,4R)-4-(4-amino-5-iodo-7H-pyrrolo[2,3-d1pyrimidin-7-yl)-2-methylpyrrolidine-1-carboxylate
[0239] tert-Butyl (2S,4S)-4-hydroxy-2-methylpyrrolidine-1-carboxylate (19.0 g) and 4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine
(13.1 g) were dissolved in THF (190 mL), and the obtained solution was then cooled
to 0°C. Thereafter, triphenylphosphine (37.2 g) and diisopropyl azodicarboxylate (28.1
mL) were added to the reaction solution, and the temperature of the mixture was then
increased to room temperature, followed by stirring for 1 hour. Thereafter, the reaction
mixture was concentrated under reduced pressure, and the obtained residue was then
purified by silica gel chromatography (hexane : ethyl acetate) to obtain the corresponding
coupling body. The obtained compound was used in the subsequent reaction without being
further purified.
[0240] The obtained coupling body, THF (114 mL) and ammonia water (114 mL) were added into
a pressure resistant tube, and the obtained mixture was then stirred at 100°C for
14 hours. Thereafter, the reaction mixture was cooled to room temperature, and was
then poured into water (285 mL). The thus obtained mixture was stirred at room temperature
for 5 hours. Thereafter, the precipitated solid was collected by filtration, was then
washed with water, and was then dried to obtain a product of interest (34.5 g).
1HNMR (CDCl3)δ: 8.27(s,1H) 7.15(s,1H) 5.55-5.73(m,2H) 5.12-5.25(m,1H) 3.86-4.18(m,2H) 3.43-3.57(m,1H)
2.59-2.69(m,1H) 1.92-2.03(m,1H) 1.48(s,9H) 1.30-1.40(m,3H)
ESI-MS m/z 444 (MH+)
Reference Example 1(2)
4-Amino-7-((3R,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidin-3-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxylic
acid
[0241] The compound of Reference Example 1(1) (28.0 g), 10% palladium carbon catalyst (720
mg), NMP (84 mL), methanol (26 mL), and triethylamine (17.6 mL) were added into a
pressure resistant tube, followed by carbon monoxide substitution, and the obtained
mixture was stirred at 100°C for 2 hours. Thereafter, the reaction mixture was cooled
to room temperature, a 2 M sodium hydroxide aqueous solution (79 mL) was then added
thereto, and the obtained mixture was then stirred at 80°C for 2 hours. Thereafter,
the reaction mixture was cooled to room temperature, was then filtrated through Celite,
and was then washed with methanol. Subsequently, methanol in the filtrate was concentrated
under reduced pressure. Water was further added, and the water layer was then washed
with tert-butyl methyl ether. A 1 M potassium hydrogen sulfate aqueous solution was
added to the water layer to adjust the pH to approximately 3. The precipitated solid
was collected by filtration, was then washed with water, and was then dried to obtain
a product of interest (23.4 g).
1HNMR (400MHz, DMSO-d6)δ: 8.14 (s, 1H) 8.08 (s, 1H) 5.16-4.93(m,1H) 4.07-3.79(m,2H)
3.61-3.45(m,1H) 2.53(m,1H) 2.33-2.02(m,1H) 1.42(s,9H) 1.29(d,J = 6.1Hz,3H) ESI-MS
m/z 362 (MH+)
< Examples >
Example 1(1)
tert-Butyl-4-amino-6-bromo-7-((3R,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidin-3-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxylate
[0242] Under a nitrogen atmosphere, the compound of Reference Example 1(2) (15.0 g) was
dissolved in chloroform (150 mL), and 2-tert-butyl-1,3-diisopropylisourea (25 mL)
was then added to the above obtained solution. The temperature of the obtained mixture
was increased to 60°C, and the mixture was then stirred for 2 hours. Thereafter, 2-tert-butyl-1,3-diisopropylisourea
(25 mL) was further added to the reaction mixture, and the thus obtained mixture was
then stirred for 2 hours. Thereafter, the reaction mixture was cooled to room temperature,
and was then concentrated under reduced pressure. To the obtained residue, tert-butyl
methyl ether was added, and the precipitated solid was collected by filtration and
was then washed with tert-butyl methyl ether. The filtrate was concentrated under
reduced pressure, and tert-butyl methyl ether was then added to the obtained residue.
The precipitated solid was collected by filtration, and was then washed with tert-butyl
methyl ether. The obtained residue was purified by silica gel chromatography (hexane
: ethyl acetate) to obtain a tert-butyl ester form. The obtained compound was used
in the subsequent halogenation reaction without being further purified.
[0243] The obtained tert-butyl ester form was dissolved in chloroform (140 mL), and N-bromosuccinimide
(11.8 g) was then added to the above obtained solution. The obtained mixture was stirred
at room temperature for 24 hours. Thereafter, to the reaction mixture, chloroform
and 10% sodium bisulfite aqueous solution were successively added, and the obtained
mixture was then extracted with chloroform. The gathered organic layer was washed
with saturated saline, was then dried over anhydrous sodium sulfate, and was then
concentrated under reduced pressure. The obtained residue was purified by silica gel
chromatography (hexane : ethyl acetate) to obtain a product of interest (13.8 g).
1HNMR (CDCl3)δ: 8.02 (s, 1H) 5.74-5.13(m,2H) 4.07-3.64(m,2H) 2.43-2.29(m,1H) 2.07-1.97(m,1H)
1.63(s,9H) 1.48(m,12H)
ESI-MS m/z 496,498 (MH+)
Example 1(2)
tert-Butyl-7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-bromo-7H-pyrrolo[2,3-d]pyrimidine-5-carboxylate
[0244] The compound of Example 1(1) (11.4 g) was dissolved in THF (57 mL), and the obtained
solution was then cooled to 0°C. Thereafter, 4 M hydrogen chloride in 1,4-dioxane
solution (114 mL) was added to the mixture, and the thus obtained mixture was then
stirred at 0°C for 10 hours. Subsequently, to the reaction mixture, a 5 M sodium hydroxide
aqueous solution (92 mL), acetonitrile (57 mL), diisopropylethylamine (20 mL), and
acryloyl chloride (2.0 mL) were added, and the obtained mixture was then stirred for
30 minutes. Thereafter, the reaction mixture was extracted with ethyl acetate, and
the gathered organic layer was washed with saturated saline, was then dried over anhydrous
sodium sulfate, and was then concentrated under reduced pressure. The obtained residue
was purified by silica gel chromatography (hexane : acetone) to obtain a product of
interest (7.72 g).
1HNMR (CDCl3)δ: 8.26-8.16(m,1H) 6.62-6.30(m,2H) 5.81-5.64(m,1H) 5.33-5.14(m,1H) 4.81-3.75(m,3H)
3.07-2.86(m,1H) 2.67-2.33(m,1H) 1.69-1.61(m,9H) 1.60-1.51(m,3H)
ESI-MS m/z 450,452 (MH+)
Example 1(3)
tert-Butyl-7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(prop-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxylate
[0245] 1.0 M Propyne in DMF solution (85.7 mL) was added to the compound of Example 1(2)
(7.72 g), acetonitrile (154 mL), triethylamine (7.2 mL), PdCl
2(PPh
3)2 (1.2 g), and copper(I) iodide (330 mg), followed by nitrogen substitution. Thereafter,
the mixture was stirred at 70°C for 4 hours. Thereafter, the reaction mixture was
cooled to room temperature, and ethyl acetate and a saturated sodium hydrogen carbonate
aqueous solution were added to the mixture. Thereafter, the obtained mixture was extracted
with ethyl acetate, and the gathered organic layer was washed with water, and then
with saturated saline. The resultant was dried over anhydrous sodium sulfate, and
was then concentrated under reduced pressure. The obtained residue was purified by
silica gel chromatography (hexane : acetone) to obtain a product of interest (4.06
g).
1HNMR (CDCl3)δ: 8.29-8.17(m,1H) 6.63-6.30(m,2H) 5.81-5.63(m,1H) 5.42-5.15(m,1H) 4.66-3.81(m,3H)
3.01-2.82(m,1H) 2.65-2.32(m,1H) 2.92-2.13(m,3H) 1.65-1.59(m,9H) 1.57-1.49(m,3H)
ESI-MS m/z 410 (MH+)
Example 1(4)
7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(prop-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-carboxylic
acid
[0246] The compound of Example 1(3) (1.52 g) was dissolved in chloroform (5 mL), and trifluoroacetic
acid (5 mL) was then added to the above obtained solution. The mixture was stirred
at room temperature for 2 hours, and the reaction mixture was then concentrated under
reduced pressure. To the residue, chloroform was added, and the obtained mixture was
concentrated under reduced pressure again. The residue was dried under reduced pressure
to obtain a product of interest (1.25 g). ESI-MS m/z 354 (MH+)
Example 1(5)
7-(R)-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-N-((R)-1-(3,5-difluorophenyl)ethyl)-6-(prop-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide
[0247] To the compound of Example 1(4) (100 mg) in DMF (1.0 mL) solution, (R)-1-(3,5-difluorophenyl)ethan-1-amine
(89.0 mg), diisopropylethylamine (0.25 mL), and HATU (215 mg) were added, and the
obtained mixture was then stirred at room temperature for 2 hours. Thereafter, to
the reaction mixture, a saturated sodium hydrogen carbonate aqueous solution was added,
and the obtained mixture was then extracted with ethyl acetate. The gathered organic
layer was washed with saturated saline, was then dried over anhydrous sodium sulfate,
and was then concentrated under reduced pressure. The obtained residue was purified
by silica gel chromatography (hexane : acetone) to obtain the title compound (60 mg).
1HNMR (DMSO-d6)6: 8.51(d,J = 7.3Hz,1H) 8.16 (s, 1H) 7.25-7.07(m,3H) 6.74-6.47(m,1H) 6.25-6.08(m,1H) 5.78-5.58(m,1H)
5.41-5.21(m,1H) 5.21-5.06(m,1H) 4.45-4.29(m,1H) 4.24-3.91(m,2H) 2.78-2.58(m,1H) 2.52-2.41(m,1H)
2.23(s,3H) 1.48(d,J= 7.1Hz,3H) 1.39(d,J= 6.1Hz,3H)
ESI-MS m/z 493 (MH+)
Example 2
7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-N-((R)-1-phenylethyl)-6-(prop-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide
[0248] The title compound was obtained in the same manner as that of Example 1, with the
exception that (R)-1-phenylethan-1-amine was used instead of (R)-1-(3,5-difluorophenyl)ethan-1-amine
in Example 1(5).
1HNMR (DMSO-d6)6: 8.35(d,J= 7.8Hz,1H) 8.17-8.13(m,1H) 7.48-7.23(m,5H) 6.76-6.46(m,1H)
6.28-6.06(m,1H) 5.81-5.58(m,1H) 5.43-5.02(m,2H) 4.42-4.28(m,1H) 4.21-3.96(m,2H) 2.74-2.59(m,1H)
2.54-2.41(m,1H) 2.17(s,3H) 1.50(d,J = 6.8Hz,3H) 1.42-1.33(m,3H)
ESI-MS m/z 457 (MH+)
Example 3
7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-N-(2-phenylpropan-2-yl)-6-(prop-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide
[0249] The title compound was obtained in the same manner as that of Example 1, with the
exception that 2-phenylpropan-2-amine was used instead of (R)-1-(3,5-difluorophenyl)ethan-1-amine
in Example 1(5).
1HNMR (DMSO-d6)6: 8.26(s,1H) 8.16-8.08(m,1H) 7.44(dd,J= 8.8,1.2Hz,2H) 7.38-7.28(m,2H)
7.21(tt,J= 7.3,1.27Hz,1H) 6.76-6.50(m,1H) 6.25-6.10(m,1H) 5.79-5.62(m,1H) 5.45-5.19(m,1H)
4.45-4.30(m,1H) 4.26-4.01(m,2H) 2.79-2.42(m,2H) 2.29-2.22(m,3H) 1.71(s,6H) 1.43-1.36(m,3H)
ESI-MS m/z 471 (MH+)
Example 4
7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-N-((R)-1-phenylpropyl)-6-(prop-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide
[0250] The title compound was obtained in the same manner as that of Example 1, with the
exception that (R)-1-phenylpropan-1-amine was used instead of (R)-1-(3,5-difluorophenyl)ethan-1-amine
in Example 1(5).
1HNMR (DMSO-d6)6: 8.35(brd,J= 8.0Hz,1H) 8.17-8.11(m,1H) 7.46-7.22(m,5H) 6.74-6.50(m,1H)
6.26-6.08(m,1H) 5.79-5.60(m,1H) 5.40-5.21(m,1H) 4.99-4.87(m,1H) 4.43-4.30(m,1H) 4.23-3.94(m,2H)
2.76-2.42(m,2H) 2.21(s,3H) 1.95-1.74(m,2H) 1.44-1.34(m,3H) 0.91(t,J= 7.3Hz,3H)
ESI-MS m/z 471 (MH+)
Example 5
7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-N-(2-(2-fluorophenyl)propan-2-yl)-6-(prop-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide
[0251] The title compound was obtained in the same manner as that of Example 1, with the
exception that 2-(2-fluorophenyl)propan-2-amine was used instead of (R)-1-(3,5-difluorophenyl)ethan-1-amine
in Example 1(5).
1HNMR (CDCl3)δ: 8.28(s,1H) 8.11(d,J = 4.4Hz,1H) 8.02(s,1H) 7.47-7.42(m,1H) 7.29-7.23(m,1H) 7.15(t,J
= 7.7Hz,1H) 7.02(ddd,J = 12.5,8.1,1.1Hz,1H) 6.58-6.35(m,2H) 5.79-5.70(m,1H) 5.30-5.19(m,1H)
4.53(t,J = 10.1Hz,0.7H) 4.38-4.25(m,1.6H) 3.92(t,J = 8.8Hz,0.7H) 2.91-2.78(m,1H) 2.70-2.60(m,0.3H)
2.54-2.43(m,0.7H) 2.28(d,J = 7.0Hz,3H) 1.88(dt,J = 10.0,5.0Hz,6H) 1.53(t,J = 6.2Hz,3H)
ESI-MS m/z 489 (MH+)
Example 6
7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-N-((R)-1-(3-chlorophenyl)ethyl)-6-(prop-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide
[0252] The title compound was obtained in the same manner as that of Example 1, with the
exception that (R)-(+)-1-(3-chlorophenyl)ethylamine hydrochloride was used instead
of (R)-1-(3,5-difluorophenyl)ethan-1-amine in Example 1(5).
1HNMR (CDCl3)δ: 8.22(d,J = 5.9Hz,1H) 7.75(d,J = 7.0Hz,1H) 7.38(s,1H) 7.35-7.27(m,3H) 6.58-6.33(m,2H)
5.78-5.66(m,1H) 5.29-5.19(m,2H) 4.56(t,J = 10.3Hz,0.7H) 4.39-4.20(m,1.6H) 3.89(t,J
= 8.8Hz,0.7H) 2.94-2.82(m,1H) 2.66-2.58(m,0.3H) 2.46(dt,J = 14.5,6.1Hz,0.7H) 2.18(d,J
= 11.0Hz,3H) 1.60(d,J = 7.0Hz,3H) 1.55-1.51(m,3H)
ESI-MS m/z 491,493 (MH+)
Example 7
7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-N-((R)-1-(2,4-difluorophenyl)ethyl)-6-(prop-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide
[0253] The title compound was obtained in the same manner as that of Example 1, with the
exception that (R)-(+)-1-(2,4-difluorophenyl)ethylamine hydrochloride was used instead
of (R)-1-(3,5-difluorophenyl)ethan-1-amine in Example 1(5).
1HNMR (CDCl3)δ: 8.20(d,J = 5.9Hz,1H) 7.98(d,J = 7.7Hz,1H) 7.37-7.31(m,1H) 6.90-6.81(m,2H) 6.58-6.35(m,2H) 5.78-5.65(m,1H) 5.44-5.37(m,1H)
5.30-5.19(m,1H) 4.56(t,J= 10.1Hz,0.7H) 4.38-4.23(m,1.6H) 3.88(t,J= 8.8Hz,0.7H) 2.94-2.83(m,1H)
2.66-2.57(m,0.3H) 2.51-2.42(m,0.7H) 2.27(d,J= 9.2Hz,3H) 1.61(d,J= 7.0Hz,3H) 1.56-1.51(m,3H)
ESI-MS m/z 493 (MH+)
Example 8
7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(prop-1-yn-1-yl)-N-((S)-2,2,2-trifluoro-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide
[0254] The title compound was obtained in the same manner as that of Example 1, with the
exception that (S)-2,2,2-trifluoro-1-phenylethan-1-amine was used instead of (R)-1-(3,5-difluorophenyl)ethan-1-amine
in Example 1(5).
1HNMR (CDCl3)δ: 8.40(d,J= 8.8Hz,1H) 8.16(s,1H) 7.44(s,5H) 6.58-6.38(m,2H) 5.92-5.84(m,1H) 5.81-5.69(m,1H)
5.29-5.19(m,1H) 4.55(t,J= 10.3Hz,0.7H) 4.41-4.24(m,1.6H) 3.91(t,J= 8.6Hz,0.7H) 2.92-2.80(m,1H)
2.70-2.61(m,0.3H) 2.54-2.46(m,0.7H) 2.35(d,J= 8.4Hz,3H) 1.54(t,J= 7.3Hz,3H)
ESI-MS m/z 511 (MH+)
Example 9
7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-(2-phenylpropan-2-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide
[0255] The title compound was obtained in the same manner as that of Example 1, with the
exceptions that cyclopropylacetylene was used instead of 1.0 M propyne in DMF solution
in Example 1(3), and that 2-phenylpropan-2-amine was used instead of (R)-1-(3,5-difluorophenyl)ethan-1-amine
in Example 1(5).
1HNMR (CDCl3)δ: 8.15(s,1H) 8.00(s,1H) 7.44(d,J= 7.7Hz,2H) 7.37(t,J = 7.7Hz,2H) 7.32-7.27(m, 1H) 6.66-6.30(m,2H) 5.81-5.69(m,1H) 5.38-5.24(m,1H) 4.48(t,J=
9.9Hz,0.7H) 4.42-4.29(m,1.6H) 4.22(t,J= 10.4Hz,0.7H) 2.77-2.68(m,1H) 2.67-2.60(m,0.3H)
2.59-2.52(m,0.7H) 1.83(s,6H) 1.60-1.52(m,4H) 1.08-1.01(m,2H) 0.92-0.88(m,2H)
ESI-MS m/z 497 (MH+)
Example 10
7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-(2,3-difluorophenyl)ethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide
[0256] The title compound was obtained in the same manner as that of Example 1, with the
exceptions that cyclopropylacetylene was used instead of 1.0 M propyne in DMF solution
in Example 1(3), and that (R)-(+)-1-(2,3-difluorophenyl)ethylamine was used instead
of (R)-1-(3,5-difluorophenyl)ethan-1-amine in Example 1(5).
1HNMR (CDCl3)δ: 8.17(d,J= 4.0Hz,1H) 8.04(d,J= 8.1Hz,1H) 7.15-7.05(m,3H) 6.58-6.36(m,2H)5.80-5.68(m,1H)
5.49-5.42(m,1H) 5.34-5.24(m,1H) 4.52(t,J = 10.1Hz,0.7H) 4.37-4.23(m,1.6H) 3.92(t,J= 8.8Hz,0.7H) 2.86-2.76(m,1H) 2.69-2.63(m,0.3H)
2.52-2.46(m,0.7H) 1.73-1.63(m,4H) 1.55(t,J= 5.3Hz,3H) 1.14-1.07(m,2H) 1.01-0.92(m,2H)
ESI-MS m/z 519 (MH+)
Example 11
Example 11(1)
tert-Butyl(2S,4R)-4-(4-amino-6-bromo-5-(((R)-1-phenylethyl)carbamoyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-methylpyrrolidine-1-carboxylate
[0257] The compound of Reference Example 1(2) (1.00 g), (R)-(+)-1-phenylethylamine (0.503
g), diisopropylethylamine (1.79 g), and N,N-dimethylformamide (10 mL) were added,
and subsequently, HATU (1.58 g) was added. The obtained mixture was stirred at room
temperature overnight. Thereafter, to the reaction mixture, ethyl acetate and a saturated
sodium hydrogen carbonate aqueous solution were added, and the obtained mixture was
then extracted with ethyl acetate. The gathered organic layer was washed with water,
and then with saturated saline. The resultant was dried over anhydrous sodium sulfate,
and was then concentrated under reduced pressure. The obtained residue was purified
by silica gel chromatography (hexane : acetone) to obtain an amide form (1.53 g).
The obtained compound was used in the subsequent reaction without being further purified.
[0258] To the amide form (1.53 g), chloroform (15 mL) was added, and the obtained mixture
was then cooled to 0°C. Thereafter, N-bromosuccinimide (0.88 g) was added to the reaction
mixture, and the obtained mixture was then stirred at 0°C for 1 hour. Thereafter,
the reaction mixture was concentrated under reduced pressure, and the obtained residue
was purified by silica gel chromatography (hexane : ethyl acetate) to obtain a product
of interest (1.39 g).
1HNMR (CDCl3)δ: 8.21 (s, 1H) 7.42-7.28(m,5H) 6.97(d,J= 7.3Hz,1H) 5.36-5.29(m,1H) 5.20-5.07(m,1H)
4.30(t,J= 10.3Hz,1H) 4.04-3.72(m,2H) 3.00-2.86(m,1H) 2.38(dt,J= 14.3,6.0Hz, 1H) 1.63(d,J=
7.0Hz,3H) 1.53-1.43(m,12H)
ESI-MS m/z 543,545 (MH+)
Example 11(2)
7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-bromo-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d1pyrimidine-5-carboxamide
[0259] To the compound of Example 11(1) (600 mg), chloroform(3 mL) was added, and the obtained
mixture was then cooled to 0°C. Thereafter, trifluoroacetic acid (4.44 g) was added
to the reaction mixture, and the thus obtained mixture was then stirred at room temperature
for 1 hour. Thereafter, the reaction mixture was concentrated under reduced pressure,
and acetonitrile (5 mL) was then added to the residue. The obtained mixture was concentrated
under reduced pressure again to obtain an amine form. The obtained compound was used
in the subsequent reaction without being further purified.
[0260] To the obtained amine form, acetonitrile (3 mL) was added, and the obtained mixture
was then cooled to 0°C. Thereafter, acryloyl chloride (99.9 mg) and diisopropylethylamine
(713 mg) were added, and the obtained mixture was then stirred at 0°C for 1 hour.
Thereafter, the reaction mixture was concentrated under reduced pressure, and the
obtained residue was purified by silica gel chromatography (ethyl acetate : methanol)
to obtain a product of interest (281 mg).
1HNMR (CDCl3)δ: 8.20(d,J= 7.3Hz,1H) 7.42-7.36(m,4H) 7.32-7.28(m,1H) 7.00-6.94(m,1H) 6.57-6.33(m,2H)
5.76-5.66(m,1H) 5.36-5.29(m, 1H) 5.14-5.08(m,1H) 4,71(t,J= 9.9Hz,0.7H) 4.42-4.23(m,1.6H)
3.83(t,J= 8.6Hz,0.7H) 3.03-2.92(m,1H) 2.60-2.57(m,0.3H) 2.44-2.40(m, 0.7H) 1.64(d,J=
6.6Hz,3H) 1.56(dd,J= 11.7,6.2Hz,3H)
ESI-MS m/z 497,499 (MH+)
Example 11(3)
7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide
[0261] The compound of Example 11(2) (65 mg), dichlorobis(triphenylphosphine)dipalladium
(9.2 mg), copper(I) iodide (5.0 mg), cyclopropylacetylene (13.0 mg), triethylamine
(39.7 mg), and N,N-dimethylformamide (1.3 mL) were added, and the inside of the reaction
system was then substituted with nitrogen. After that, the mixture was stirred at
70°C for 2.5 hours. Thereafter, to the reaction mixture, ethyl acetate and a saturated
ammonium chloride aqueous solution were added, and the obtained mixture was then extracted
with ethyl acetate. The gathered organic layer was washed with water, and then with
saturated saline. The resultant was dried over anhydrous sodium sulfate, and was then
concentrated under reduced pressure. The obtained residue was purified by silica gel
chromatography (chloroform : methanol) to obtain a product of interest (50 mg).
1HNMR (CDCl3)δ: 8.22(d,J = 5.1Hz,1H) 7.82(d,J= 7.3Hz,1H) 7.43-7.35(m,4H) 7.30(t,J= 6.8Hz,1H) 6.58-6.34(m,2H)
5.77-5.66(m,1H) 5.35-5.20(m,2H) 4.54(t,J= 10.1Hz,0.7H) 4.35-4.25(m,1.6H) 3.88(t,J=
8.8Hz,0.7H) 2.90-2.78(m,1H) 2.65-2.56(m,0.3H) 2.49-2.40(m,0.7H) 1.63 (d,J = 7.0Hz,3H)
1.56-1.45(m,4H) 1.03-0.91(m,2H) 0.84-0.69(m,2H)
ESI-MS m/z 483 (MH+)
Example 12
7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(3,3-dimethylbut-1-yn-1-yl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide
[0262] The title compound was obtained in the same manner as that of Example 11, with the
exception that 3,3-dimethyl-1-butyne was used instead of cyclopropylacetylene in Example
11(3).
1HNMR (CDCl3)δ: 8.22(d,J= 5.9Hz,1H) 7.75(d,J= 7.7Hz,1H) 7.38(dt,J = 15.5,7.1Hz,4H) 7.31-7.25(m,1H) 6.57-6.34(m,2H) 5.77-5.65(m,1H) 5.44-5.35(m,1H)
5.33-5.15(m,1H) 4.63(t,J= 10.1Hz,0.7H) 4.40-4.20(m,1.6H) 3.89(t,J= 8.8Hz,0.7H) 2.90-2.76(m,1H)
2.65-2.55(m,0.3H) 2.49-2.40(m,0.7H) 1.85(s,1H) 1.64(d,J= 7.0Hz,3H) 1.55(d,J= 5.9Hz,3H)
1.26(s,9H)
ESI-MS m/z 499 (MH+)
Example 13
7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(3-methoxy-3-methylbut-1-yn-1-yl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,2-d]pyrimidine-5-carboxamide
[0263] The title compound was obtained in the same manner as that of Example 11, with the
exception that 3-methoxy-3-methyl-1-butyne was used instead of cyclopropylacetylene
in Example 11(3).
1HNMR (CDCl3)δ: 8.17 (s, 1H) 7.61(d,J= 7.7Hz,1H) 7.43-7.35(m,4H) 7.30(d,J = 7.0Hz,1H) 6.57-6.33(m,2H) 5.81-5.68(m,1H) 5.43-5.33(m,1H) 5.29-5.12(m,1H) 4.59(t,J=
10.1Hz,0.7H) 4.38-4.22(m,1.6H) 3.92(t,J= 8.6Hz,0.7H) 3.30(s,3H) 2.86-2.72(m,1H) 2.70-2.60(m,1.3H)
2.52-2.44(m,0.7H) 1.64(d,J= 7.0Hz,3H) 1.55(t,J= 5.5Hz,3H) 1.46(d,J= 2.2Hz,6H)
ESI-MS m/z 515 (MH+)
Example 14
7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(but-1-yn-1-yl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide
[0264] The title compound was obtained in the same manner as that of Example 11, with the
exception that 1-trimethylsilyl-1-butyne and tetra-n-butylammonium fluoride were used
instead of cyclopropylacetylene in Example 11(3).
1HNMR (CDCl3)δ: 8.26-8.25(m,1H) 7.79(d,J = 7.3Hz,1H) 7.42-7.36(m,4H) 7.32-7.30(m,1H) 6.57-6.37(m,2H)
5.76-5.66(m,1H) 5.33-5.20(m,2H) 4.57(t,J = 10.3Hz,0.7H) 4.36-4.22(m,1.6H) 3.88(t,J= 8.8Hz,0.7H) 2.92-2.81(m,1H) 2.65-2.57(m,0.3H)
2.48-2.38(m,2.7H) 1.63 (d,J = 7.0Hz,3H) 1.54-1.51(m,3H) 1.17-1.12(m,3H)
ESI-MS m/z 471 (MH+)
Example 15
7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-N-(2-(2-fluorophenyl)propan-2-yl)-6-(3-methylbut-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide
[0265] The title compound was obtained in the same manner as that of Example 11, with the
exceptions that 2-(2-fluorophenyl)propan-2-amine was used instead of (R)-(+)-1-phenylethylamine
in Example 11(1), and that 3-methyl-1-butyne was used instead of cyclopropylacetylene
in Example 11(3).
1HNMR (CDCl3)δ: 7.92 (s, 1H) 7.44(t,J = 7.9Hz,1H) 7.30-7.23(m,1H) 7.14(t,J = 7.5Hz,1H) 7.02(dd,J= 12.6,8.2Hz,1H) 6.58-6.35(m,2H) 5.80-5.69(m,1H) 5.33-5.16(m,1H)
4.58(t,J= 9.9Hz,0.7H) 4.38-4.23(m,1.6H) 3.91(t,J= 8.4Hz,0.7H) 3.03-2.93(m,1H) 2.89-2.75(m,1H)
2.69-2.60(m,0.3H) 2.53-2.43(m,0.7H) 1.88(s,6H) 1.55(d,J = 5.1Hz,3H) 1.36(d,J = 6.6Hz,6H)
ESI-MS m/z 517 (MH+)
Example 16
Example 16(1)
tert-Butyl (2R,4S)-4-(benzyloxy)-2-((tosyloxy)methyl)pyrrolidine-1-carboxylate
[0266] tert-Butyl (2R,4S)-4-(benzyloxy)-2-(hydroxymethyl)pyrrolidine-1-carboxylate (2.0
g) was dissolved in methylene chloride (20 mL), and the obtained solution was then
cooled to 0°C. Thereafter, 1,4-diazabicyclo[2.2.2]octane (2.2 g) and tosylate chloride
(1.9 g) were added to the reaction solution, and the temperature of the mixture was
then increased to room temperature. The mixture was stirred for 4 hours. Thereafter,
a saturated sodium hydrogen carbonate aqueous solution was added to the reaction mixture,
and the obtained mixture was then extracted with ethyl acetate. The gathered organic
layer was washed with saturated saline, was then dried over anhydrous sodium sulfate,
and was then concentrated under reduced pressure. The obtained residue was purified
by silica gel chromatography (hexane : ethyl acetate) to obtain a product of interest
(4.32 g).
1HNMR (CDCl3)δ: 7.78(d,J = 8.1Hz,2H), 7.42-7.29(m,7H), 4.57-4.41(m,2H), 4.39-3.96(m,4H), 3.61-3.20(m,2H),2.46(s,3H),
2.27-2.02(m,2H), 1.48-1.31(m,9H)
ESI-MS m/z 462 (MH+)
Example 16(2)
tert-Butyl (2S,4S)-4-(benzyloxy)-2-ethylpyrrolidine-1-carboxylate
[0267] Under a nitrogen atmosphere, copper iodide (2.04 g) was suspended in diethyl ether
(12 mL), and the obtained suspension was then cooled to 0°C. Thereafter, 1.04 M methyl
lithium in diethyl ether solution (0.36 mL) was added, and the obtained mixture was
then stirred at 0°C for 30 minutes. Subsequently, the compound of Example 16(1) (1.98
g) in methylene chloride (4.0 mL) solution was added to the reaction mixture, and
the temperature of the obtained mixture was then increased to room temperature. The
mixture was stirred for 1 hour. Thereafter, the reaction mixture was cooled to 0°C,
and a saturated ammonium chloride aqueous solution was then added to the reaction
mixture. The thus obtained mixture was extracted with ethyl acetate. The gathered
organic layer was washed with saturated saline, was then dried over anhydrous sodium
sulfate, and was then concentrated under reduced pressure. The obtained residue was
purified by silica gel chromatography (hexane : ethyl acetate) to obtain a product
of interest (707 mg).
1HNMR (CDCl3)δ 7.42-7.25(m,5H), 4.66-4.40(m,2H), 4.17-4.03(m,1H), 4.00-3.26(m,3H), 2.24-2.09(m,1H),
1.96-1.71(m,2H), 1.48(s,9H),1.45- 1.31(m,1H), 0.86(t,J = 7.4Hz,3H)
ESI-MS m/z 306 (MH+)
Example 16(3)
tert-Butyl (2S,4S)-2-ethyl-4-hydroxypyrrolidine-1-carboxylate
[0268] The compound of Example 16(2) (1.06 g) and a 10% palladium hydroxide carbon catalyst
(160 mg) were suspended in ethanol (11 mL) and THF (11 mL), followed by hydrogen substitution,
and the resultant was then stirred at room temperature for 20 hours. Thereafter, the
reaction mixture was filtrated through Celite, and was then washed with ethanol, and
the filtrate was then concentrated under reduced pressure. The obtained residue was
purified by silica gel chromatography (hexane : ethyl acetate) to obtain a product
of interest (709 mg).
1HNMR (CDCl
3)δ 4.46-4.36(m,1H), 4.02-3.81(m,1H), 3.71-3.35(m,2H), 2.15-1.99(m,1H), 1.95-1.72(m,2H),
1.49(s,9H), 1.46-1.35(m,1H), 0.86(t,J = 7.5Hz,3H) ESI-MS m/z 216 (MH
+)
Example 16(4)
tert-Butyl (2S,4R)-4-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-ethylpyrrolidine-1-carboxylate
[0269] The compound of Example 16(3) (709 mg) and 4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine
(1.11 g) were dissolved in THF (7.1 mL), and the obtained solution was then cooled
to 0°C. Thereafter, triphenylphosphine (1.3 g) and diisopropyl azodicarboxylate (1.00
mL) were added, and the temperature of the obtained mixture was then increased to
room temperature, followed by stirring the mixture for 1 hour. Thereafter, the reaction
mixture was concentrated under reduced pressure, and the obtained residue was then
purified by silica gel chromatography (hexane : ethyl acetate) to obtain the corresponding
coupling body. The obtained compound was used in the subsequent reaction without being
further purified. Into a pressure resistant tube, the obtained coupling body, THF
(5.4 mL), and ammonia water (5.4 mL) were added, and the obtained mixture was then
stirred at 100°C for 14 hours. Thereafter, the reaction mixture was cooled to room
temperature, and was then poured into water (12.8 mL), and the mixed solution was
then extracted with ethyl acetate. The gathered organic layer was washed with saturated
saline, was then dried over anhydrous sodium sulfate, and was then concentrated under
reduced pressure. The obtained residue was purified by silica gel chromatography (hexane
: acetone) to obtain a product of interest (797 mg).
1HNMR (CDCl3)δ 8.29(s,1H), 7.14(s,1H), 5.67(br s,2H), 5.32-5.09(m,1H), 4.24-4.08(m,1H), 3.95-3.79(m,1H),
3.46(dd,J = 9.3,11.0Hz,1H), 2.70-2.55(m,1H), 2.06-1.95(m,1H), 1.59-1.51(m,2H), 1.49(s,9H),
0.91(t,J = 7.5Hz,3H)
ESI-MS m/z 458 (MH+)
Example 16(5)
tert-Butyl (2S,4R)-4-(4-amino-6-bromo-5-(((R)-1-phenylethyl)carbamoyl)-7H-pyrrolo
[2,3-d]pyrimidin-7-yl)-2-ethylpyrrolidine-1-carboxylate
[0270] The compound of Example 16(4) (797 mg), dichlorobis(triphenylphosphine)dipalladium
(25 mg), and (R)-(+)-1-phenylethylamine (0.55 mL) were suspended in DMF (8.0 mL),
followed by carbon monoxide substitution, and the resultant was then stirred at 80°C
for 2 hours. Thereafter, the reaction mixture was cooled to room temperature, water
was then added thereto, and the obtained mixture was then extracted with ethyl acetate.
The gathered organic layer was washed with saturated saline, was then dried over anhydrous
sodium sulfate, and was then concentrated under reduced pressure. The obtained residue
was purified by silica gel chromatography (hexane : acetone) to obtain the corresponding
amide form. The obtained compound was used in the subsequent reaction without being
further purified. The obtained amide form was dissolved in acetonitrile (8.2 mL),
and the obtained solution was then cooled to - 10°C. Thereafter, N-bromosuccinimide
(457 mg) in acetonitrile (8.2 mL) solution was slowly added dropwise to the solution,
and the reaction mixture was then stirred for 30 minutes. Thereafter, to the reaction
mixture, a sodium sulfite aqueous solution and a sodium hydrogen carbonate aqueous
solution were added, and the obtained mixture was then extracted with ethyl acetate.
The gathered organic layer was washed with saturated saline, was then dried over anhydrous
sodium sulfate, and was then concentrated under reduced pressure. The obtained residue
was purified by silica gel chromatography (hexane : acetone) to obtain a product of
interest (650 mg).
1HNMR (CDCl3)δ 8.23(s,1H), 7.49-7.29(m,5H), 6.98(d,J = 7.4Hz,1H), 5.41-5.28(m,1H), 5.24-5.04(m,1H),
4.38-4.22(m,1H), 4.07-3.68(m,1H), 3.19-2.83(m,1H), 2.43-2.29(m,1H), 2.25-1.67(m,3H),
1.66(d,J = 6.9Hz,3H), 1.51(s,9H), 0.98(t,J = 7.4Hz,3H)
ESI-MS m/z 557,559 (MH+)
Example 16(6)
7-((3R,5S)-1-acryloyl-5-ethylpyrrolidin-3-yl)-4-amino-6-bromo-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide
[0271] To the compound of Example 16(5) (650 mg), acetonitrile (9.7 mL) was added, and the
obtained mixture was then cooled to 0°C. Thereafter, sodium iodide (1.05 g) and trimethylsilyl
chloride (0.89 mL) were added, and the obtained mixture was then stirred at 0°C for
1 hour. Thereafter, to the reaction mixture, ethanol (9.7 mL), isopropylethylamine
(2.0 mL), and acrylic acid anhydride (0.16 mL) were successively added, and the obtained
mixture was then stirred at 0°C for 30 minutes. Thereafter, to the reaction mixture,
ammonia water and water were added, and the obtained mixture was then extracted with
ethyl acetate. The gathered organic layer was washed with saturated saline, was then
dried over anhydrous sodium sulfate, and was then concentrated under reduced pressure.
The obtained residue was purified by silica gel chromatography (hexane : acetone)
to obtain a product of interest (256 mg).
1HNMR (CDCl3)δ 8.27-8.16(m,1H), 7.47-7.29(m,5H), 6.98(d,J = 7.3Hz,1H), 6.61-6.29(m,2H), 5.84-5.63(m,1H),
5.43-5.26(m,1H), 5.22-5.01(m,1H), 4.80-3.82(m,3H), 3.23-2.92(m,1H), 2.58-2.30(m,1H),
2.22-1.79(m,2H),1.66(d,J = 7.0Hz,3H),1.07-0.96(m,3H)
ESI-MS m/z 511,513 (MH+)
Example 16(7)
7-((3R,5S)-1-acryloyl-5-ethylpyrrolidin-3-yl)-4-amino-N-((R)-1-phenylethyl)-6-(prop-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide
[0272] 1.0 M Propyne in DMF solution (0.70 mL) was added to the compound of Example 16(6)
(120 mg), acetonitrile (1.2 mL), triethylamine (0.10 mL), PdCl
2(PPh
3)
2 (8.2 mg), and copper(I) iodide (0.4 mg), followed by nitrogen substitution, and the
mixture was then stirred at 60°C for 2 hours. Thereafter, the reaction mixture was
cooled to room temperature, and ethyl acetate and a saturated ammonium chloride aqueous
solution were added to the mixture. The thus obtained mixture was extracted with ethyl
acetate, and the gathered organic layer was washed with water, and then with saturated
saline. The resultant was dried over anhydrous sodium sulfate, and was then concentrated
under reduced pressure. The obtained residue was purified by silica gel chromatography
(ethyl acetate : methanol) to obtain a product of interest (102 mg).
1HNMR (CDCl3)δ: 8.26(s,1H), 7.79(br d,J = 7.0Hz,1H), 7.46-7.30(m,5H), 6.58-6.31(m,2H), 5.80-5.65(m,1H),
5.33-5.15(m,2H), 4.59-3.85(m,3H), 3.03-2.33(m,2H), 2.25-1.70(m,5H), 1.65(d,J = 6.8Hz,6H),
1.09-0.91(m,3H)
ESI-MS m/z 471 (MH+)
Example 17
7-((3R,5S)-1-acryloyl-5-ethylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide
[0273] The title compound was obtained in the same manner as that of Example 16, with the
exception that cyclopropylacetylene was used instead of 1.0 M propyne in DMF solution
in Example 16(7).
1HNMR (CDCl3)δ: 8.31-8.16(m,1H), 7.84(d,J = 7.4Hz,1H), 7.46-7.30(m,5H), 6.64-6.32(m,2H), 5.82-5.67(m,1H),
5.39-5.17(m,2H), 4.67-3.81(m,3H), 3.02-2.80(m,1H), 2.62-1.71(m,3H), 1.65(d,J = 6.9Hz,3H),
1.58-1.47(m,1H), 1.06-0.92(m,5H), 0.85-0.70(m,2H)
ESI-MS m/z 497 (MH+)
Example 18
7-((3R,5R)-1-acryloyl-5-(methoxymethyl)pyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide
[0274] The title compound was obtained in the same manner as that of Example 16, with the
exceptions that tert-butyl (2R,4S)-4-hydroxy-2-(methoxymethyl)pyrrolidine-1 -carboxylate
was used instead of the compound of Example 16(3) in Example 16(4), and that cyclopropylacetylene
was used instead of 1.0 M propyne in DMF solution in Example 16(7).
1HNMR (CDCl3)δ: 8.29-8.22(m,1H), 7.86-7.80(m,1H), 7.36-7.44(m,4H), 7.34-7.28(m,1H), 6.48-6.37(m,2H),
5.78-5.69(m,1H), 5.29-5.15(m,2H), 4.55-4.30(m,2H), 3.96-3.65(m,3H), 3.42(s,3H), 3.18-3.06(m,0.3H),
2.90-2.80(m,0.3H), 2.64-2.58(m,0.3H), 2.47-2.35(m,0.7H),1.64(d,3H,J = 6.9Hz), 1.58-1.47(m,1H),
1.04-0.94(m,2H), 0.87-0.69(m,2H)
ESI-MS m/z 513 (MH+)
Example 19
7-((3R,5R)-1-acryloyl-5-(ethoxymethyl)pyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide
[0275] The title compound was obtained in the same manner as that of Example 16, with the
exceptions that tert-butyl (2R,4S)-2-(ethoxymethyl)-4-hydroxypyrrolidine-1 -carboxamide
was used instead of the compound of Example 16(3) in Example 16(4), and that cyclopropylacetylene
was used instead of 1.0 M propyne in DMF solution in Example 16(7).
1HNMR (CDCl3)δ: 8.28-8.18(m,1H),7.84(br d,J = 7.0Hz,1H), 7.47-7.29(m,5H), 6.82-6.35(m,2H), 5.79-5.68(m,1H),
5.40-5.14(m,2H), 4.63-3.53(m,7H), 3.20-2.79(m,1H), 2.69-2.40(m,1H), 1.67-1.63(m,3H),
1.59-1.47(m,1H),1.22(t,J = 7.0Hz,3H), 1.05-0.92(m,2H), 0.87-0.72(m,2H)
ESI-MS m/z 527 (MH+)
Comparative Example 1
4-Amino-N-(4-(methoxymethyl)phenyl)-7-(1-methylcyclopropyl)-6-(prop-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide
[0276] The title compound was obtained by the method described in Example 95 of International
Publication No.
WO 2017/146116.
ESI-MS m/z 390 (MH+)
Comparative Example 2
1-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-N-(4-(2-(dimethylamino)-2-oxoethyl)-2,3-dimethylphenyl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide
[0277] The title compound was obtained by the method described in Example 79 of International
Publication No.
WO 2017/038838.
ESI-MS m/z 505 (MH+)
Comparative Example 3
7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-N-(cyclohexylmethyl)-6-(prop-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide
[0278] The title compound was obtained in the same manner as that of Example 1, with the
exception that cyclohexylmethanamine was used instead of (R)-1-(3,5-difluorophenyl)ethan-1-amine
in Example 1(5).
1HNMR (DMSO-d6)δ:8.68-8.31(m,1H) 8.20-8.10(m,1H) 8.09-7.97(m,1H) 7.59-7.20(m,1H) 6.74-6.49(m,1H)
6.25-6.09(m,1H) 5.78-5.60(m,1H) 5.40-5.20(m,1H) 4.44-4.29(m,1H) 4.23-3.92(m,2H) 3.25-3.12(m,2H)
2.76-2.40(m,2H) 2.25(s,3H) 1.81-1.45(m,5H) 1.43-1.34(m,3H) 1.30-0.90(m,6H)
ESI-MS m/z 449 (MH+)
Comparative Example 4
7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-N-(2-methylbenzyl)-6-(prop-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide
[0279] The title compound was obtained in the same manner as that of Example 1, with the
exception that o-tolylmethanamine was used instead of (R)-1-(3,5-difluorophenyl)ethan-1-amine
in Example 1(5).
1HNMR (DMSO-d6)δ:8.37-8.27(m,1H) 8.19-8.09(m,1H) 7.39-7.30(m,1H) 7.26-7.11(m,4H) 6.68-6.48(m,1H)
6.24-6.07(m,1H) 5.80-5.60(m,1H) 5.36-5.17(m,1H) 4.52(d,J= 5.7Hz,2H) 4.42-4.28(m,1H)
4.22-3.92(m,2H) 2.73-2.42(m,2H) 2.33(s,3H) 2.02(s,3H) 1.43-1.32(m,3H)
ESI-MS m/z 457 (MH+)
Comparative Example 5
7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-N-methyl-N-((R)-1-phenylethyl)-6-(prop-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide
[0280] The title compound was obtained in the same manner as that of Example 1, with the
exception that (R)-N-methyl-1-phenylethan-1-amine was used instead of (R)-1-(3,5-difluorophenyl)ethan-1-amine
in Example 1(5).
1HNMR (CDCl3)δ: 8.23(d,J = 5.9Hz,1H) 7.50-7.28(m,4H) 7.09-6.88(m,1H) 6.57-6.34(m,2H) 5.79-5.64(m,1H) 5.22(t,J=
9.3Hz,1H) 4.48(t,J= 9.7Hz,0.6H) 4.39-4.20(m,1.9H) 3.90(t,J= 8.6Hz,0.5H) 2.85(s,4H)
2.66-2.63(m,0.4H) 2.51-2.44(m,0.6H) 2.07(s,2H) 1.66(d,J= 4.8Hz,3H) 1.52(d,J= 5.9Hz,3H)
ESI-MS m/z 471 (MH+)
Comparative Example 6
Comparative Example 6(1)
tert-Butyl (2S,4R)-4-(4-amino-5-(((R)-1-phenylethyl)carbamoyl)-6-(prop-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-methylpyrrolidine-1-carboxylate
[0281] 1.0 M Propyne in DMF solution(2.1 mL) was added to the compound of Example 11(1)
(230 mg), acetonitrile (4.6 mL), triethylamine (0.29 mL), PdCl
2(PPh
3)
2 (5.9 mg), and copper(I) iodide (1.6 mg), followed by nitrogen substitution, and the
obtained mixture was then stirred at 70°C for 1 hour. Thereafter, the reaction mixture
was cooled to room temperature, and ethyl acetate and a saturated sodium hydrogen
carbonate aqueous solution were then added to the mixture. The thus obtained mixture
was extracted with ethyl acetate, and the gathered organic layer was washed with water
and then with saturated saline. The resultant was dried over anhydrous sodium sulfate,
and was then concentrated under reduced pressure. The obtained residue was purified
by silica gel chromatography (hexane : ethyl acetate) to obtain a product of interest
(193 mg).
1HNMR (CDCl3)δ: 8.23 (s, 1H) 7.79(d,J = 6.8Hz,1H)7.46-7.27(m,5H) 5.40-5.17(m,2H) 4.28-3.64(m,3H) 2.85-2.68(m,1H) 2.46-2.36(m,1H)
2.15-1.97(m,3H) 1.62(d,J= 6.8Hz,3H) 1.56-1.32(m,12H)
ESI-MS m/z 503 (MH+)
Comparative Example 6(2)
4-Amino-7-((3R,5S)-5-methylpyrrolidin-3-yl)-N-((R)-1-phenylethyl)-6-(prop-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide
hydrochloride
[0282] To the compound of Comparative Example 6(1) (530 mg), 4 M hydrochloric acid in 1,4-dioxane
solution (5 mL) was added, and the obtained mixture was then stirred at room temperature
for 2 hours. Thereafter, the reaction mixture was concentrated under reduced pressure
to obtain a product of interest (420 mg).
ESI-MS m/z 403 (MH+)
Comparative Example 6(3)
4-Amino-7-((3R,5S)-1-((E)-but-2-enoyl)-5-methylpyrrolidin-3-yl)-N-((R)-1-phenylethyl)-6-(prop-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide
[0283] To the compound of Comparative Example 6(2) (18 mg), acetonitrile (0.5 mL) was added,
and the obtained mixture was then cooled to 0°C. Thereafter, acryloyl chloride (0.004
mL) and diisopropylethylamine (0.036 mL) were added to the reaction mixture, and the
thus obtained mixture was then stirred at 0°C for 1 hour. Thereafter, the reaction
mixture was concentrated under reduced pressure, and was then subjected to reverse
phase preparative HPLC (water : acetonitrile (0.1% formic acid)) to obtain a product
of interest (8.7 mg).
1HNMR (CDCl3)δ: 8.31 (s, 1H) 8.14(d,J= 6.2Hz,1H)7.77(d,J = 7.0Hz, 1H) 7.39-7.36(m,4H) 7.33-7.31(m,1H) 7.03-6.90(m,1H) 6.06(dd,J = 14.3Hz,1H) 5.28-5.17(m,2H) 4.48(t,J = 10.1Hz,1H) 4.35-4.20(m,2H) 3.88(t,8.8Hz,1H) 2.84-2.76(m,1H) 2.64-2.40(m,1H) 2.05(d,J=
10.6Hz,3H) 1.92(d,J= 6.6Hz,1H) 1.85(d,J = 7.0Hz,2H) 1.62(d,J= 7.0Hz,3H) 1.55(dd,J=
9.0,5.7Hz,3H)
ESI-MS m/z 471 (MH+)
[0284] The compounds synthesized in the above-described Examples and Comparative Examples
are shown below.
Test Example 1 Measurement of phosphorylation activity inhibitory effect (in vitro) on wild-type and mutant EGFR
[0285] The measurement of the
in vitro inhibitory activity of a compound against wild-type and mutant EGFR was outsourced
to Carna Biosciences, Inc. (Kinase Profiling Book, https://www.carnabio.com/japanese/product/search.cgi?mode=profiling).
[0286] Specifically, first, the compound of the present invention was diluted stepwise with
dimethyl sulfoxide (DMSO). Subsequently, the EGFR protein, the substrate peptide (Srctide,
final concentration: 1 µM), magnesium chloride (final concentration: 5 mM), manganese
chloride (final concentration: 1 mM), ATP (final concentration: near Km of each EGFR),
and the compound of the present invention in DMSO solution (final concentration of
DMSO: 1%) were added to a buffer for the kinase reaction (20 mM HEPES (pH 7.5), 1
mM dithiothreitol, and 0.01% Triton X-100), and the obtained mixture was then incubated
at room temperature for 1 hour, so that the kinase reaction was carried out. To the
reaction solution, Termination Buffer was added, so as to terminate the kinase reaction.
Finally, using LabChip(TM) EZ Reader II (PerkinElmer), an unphosphorylated substrate
peptide (S) and a phosphorylated peptide (P) were separated and detected according
to microchannel capillary electrophoresis. From the peak heights of S and P, the amount
of the phosphorylation reaction was obtained, and the concentration of the compound
capable of inhibiting the phosphorylation reaction by 50% was defined as an IC50 value
(nM). The results are shown in the following table.
[Table 3]
|
IC50 (nmol/L) |
kinases |
Example compound 11 |
EGFR |
0.47 |
EGFR [d746-750] |
0.25 |
EGFR [d746-750/T790M] |
6.8 |
EGFR [L858R] |
0.36 |
EGFR [T790M/L858R] |
6.6 |
EGFR [D770_N771insNPG] |
0.55 |
[0287] From the above results, it was found that the compound of the present invention has
excellent inhibitory activity against wild-type and mutant EGFR.
Test Example 2 Measurement of growth inhibitory activity against EGFR overexpressing
cell line and exon 20 insertion mutant EGFR expressing cell line
[0288] Growth inhibitory activity against an EGFR overexpressing cell line and an exon 20
insertion mutant EGFR expressing cell line was evaluated using MCF10A_EGFR cells,
MCF10A_EGFR/V769_D770insASV cells, MCF10A_EGFR/D770_N771insSVD cells, and MCF10A_EGFR/H773_V774insNPH
cells (Fukushima Medical University) which were cells obtained by introducing the
EGFR gene (WT, D769_N770insASV mutant, D770_N771insSVD mutant, H773_V774insNPH mutant)
into MDA-MB-468 cells (ATCC) as an EGFR overexpressing human breast cancer cell line,
NCI-H1975 cells (ATCC) as L858R and T790M mutant EGFR-positive human lung cancer cells,
and MCF10A cells as human normal mammary gland cells.
[0289] The MDA-MB-468 cells were suspended in Leibovitz's L-15 medium containing 10% inactivated
fetal bovine serum. The NCI-H1975 cells were suspended in RPMI-1640 medium containing
10% inactivated fetal bovine serum. The MCF10A_EGFR cells, the MCF10A_EGFR/V769_D770insASV
cells, the MCF10A_EGFR/D770_N771insSVD cells, or the MCF10A_EGFR/H773_V774insNPH cells
were suspended in DMEM/Ham's F-12 medium (containing L-glutamine, phenol red, HEPES,
and sodium pyruvate) containing 10 µg/mL insulin, 500 ng/mL hydrocortisone, 5 µmol/L
forskolin, and 5% inactivated horse serum in terms of their final concentrations.
Each cell suspension was seeded in each well of a 96-well flat-bottom plate such that
the number of cells per well was 500, and was then cultured in a carbon dioxide gas-free
culture vessel for the MDA-MB-468 cells and in a 5% carbon dioxide gas-containing
culture vessel for the other cells at 37°C for 1 day. The compound of the present
invention was prepared at 1 mM in DMSO and then diluted 1/200 with a medium to prepare
a 5 µM solution. Thereafter, the compound of the present invention in the DMSO solution
was diluted with the medium used in the suspension of the cells, and the obtained
solution was then added to each well so that the final concentration of the highest
concentration of the test compound was 1000 nM. The obtained mixture was further cultured
in a carbon dioxide gas-free culture vessel for the MDA-MB-468 cells and in the 5%
carbon dioxide gas-containing culture vessel for the other cells at 37°C for 3 days.
[0290] The cells at the start of the culture (day 0) and after the culture (day 3) were
counted using CellTiter-Glo(R) 2.0 Reagent (Promega Corp.) according to the protocol
recommended by the manufacturer. The growth inhibition percentage was calculated according
to the equation given below to determine the 50% inhibition concentration (GI50 (nM))
of the test compound. The results are shown in Table 4.
- 1) In the case of Tday3 ≥ Cday0
T: Emission intensity from the well to which the test compound was added
C: Emission intensity from the well to which the test compound was not added
day0: Day on which the test compound was added
day3: Evaluation day
- 2) In the case of Tday3 < Cday0
T: Emission intensity from the well to which the test compound was added
C: Emission intensity from the well to which the test compound was not added
day0: Day on which the test compound was added
day3: Evaluation day
[Table 4]
|
Cell growth inhibitory activity GI 50 value (nM) |
MDA-MB-468 |
NCI-H1975 |
MCF10A EGFR cell (EGFR-WT) |
MCF10A EGFR/ V769_D770insASV cell |
MCF10A EGFR/ D770_N771insSVD cell |
MCF10A_EGFR/ H773_V774insNPH cell |
Example compound 2 |
81 |
77 |
55 |
6.7 |
8.2 |
16 |
Example compound 11 |
99 |
87 |
81 |
7.3 |
10 |
18 |
Example compound 12 |
221 |
153 |
206 |
24 |
27 |
50 |
[0291] From the above results, it was found that the compound group of the present invention
also has excellent cell growth inhibitory activity against the wild-type EGFR overexpressing
line MDA-MB-468 cells, the MCF10A_EGFR cells expressing the introduced wild-type EGFR
gene, the L858R and T790M mutant EGFR-positive cells NCI-H1975 cells, and the exon
20 insertion mutant EGFR expressing cell lines (MCF10A_EGFR/V769_D770insASV cells,
MCF10A_EGFR/D770_N771insSVD cells, and MCF10A_EGFR/H773_V774insNPH cells).
Test Example 3 Measurement of growth inhibitory activity against exon 20 insertion
mutant EGFR expressing cell line
[0292] Growth inhibitory activity against exon 20 insertion mutant EGFR was evaluated using
H1975-EGFRinsSVD cells obtained by genetically modifying NCI-H1975 cells so as to
express D770_N771insSVD mutant EGFR and to knock out endogenous EGFR (T790M/L858R),
and LXF 2478 cells (Charles River Laboratories, Inc.) of V769_D770insASV mutant EGFR-positive
human lung cancer patient-derived tumor.
[0293] The H1975-EGFRinsSVD cells were prepared by introducing PB-CMV-MCS-EF1-RFP+Puro vector
encoding D770_N771insSVD (insSVD), together with Super PiggyBacTransposase expression
vector, to NCI-H1975 cells by electroporation with Amaxa(R) Cell Line Nucleofector(R)
Kit R, then selecting cells using puromycin (Sigma-Aldrich Co. LLC.), then introducing
thereto XTN(R) TALENs Site-Specific Nucleases (Transposagen Bio) by electroporation
with Amaxa(R) Cell Line Nucleofector(R) Kit R, and selecting, by sequencing, cells
in which endogenous EGFR (T790M/L858R) was knocked out.
[0294] Upon evaluation of cell growth inhibitory effect, the cells of each line were suspended
in RPMI-1640 medium. The cell suspension was seeded in each well of a 96-well flat-bottom
plate such that the number of cells per well was 3,000, and was then cultured in a
5% carbon dioxide gas-containing culture vessel at 37°C for 1 day. The compound of
the present invention or a Comparative Example compound was dissolved at 1 mM in DMSO
and then added to each well using Tecan D300e digital dispenser (Tecan Trading AG)
such that the final concentration of the highest concentration of the test compound
was 1000 nM and the common ratio was 3. The cells were cultured in a 5% carbon dioxide
gas-containing culture vessel at 37°C for 3 days. The cells at the start of the culture
(day 0) and after the culture (day 3) were counted using CellTiter-Glo(R) 2.0 Reagent
(Promega Corp.) according to the protocol recommended by the manufacturer. The growth
inhibition percentage was calculated according to the equation given below to determine
the 50% inhibition concentration (GI50 (nM)) of the test compound. The results are
shown in Table 5.
- 1) In the case of Tday3 ≥ Cday0
T: Emission intensity from the well to which the test compound was added
C: Emission intensity from the well to which the test compound was not added
day0: Day on which the test compound was added
day3: Evaluation day
- 2) In the case of Tday3 < Cday0
T: Emission intensity from the well to which the test compound was added
C: Emission intensity from the well to which the test compound was not added
day0: Day on which the test compound was added
day3: Evaluation day
[Table 5]
|
Cell growth inhibitory activity GI 50 value (nM) |
H1975-EGFRinsSVD cell |
LXF 2478 cell |
Example compound 1 |
44 |
29 |
Example compound 2 |
9.5 |
11 |
Example compound 3 |
77 |
24 |
Example compound 4 |
13 |
15 |
Example compound 5 |
33 |
24 |
Example compound 6 |
29 |
29 |
Example compound 7 |
33 |
33 |
Example compound 8 |
25 |
26 |
Example compound 9 |
65 |
34 |
Example compound 10 |
39 |
40 |
Example compound 11 |
17 |
19 |
Example compound 12 |
52 |
48 |
Example compound 13 |
48 |
46 |
Example compound 14 |
30 |
27 |
Example compound 15 |
101 |
58 |
Comparative Example 1 |
> 1000 |
> 1000 |
Comparative Example 2 |
> 1000 |
> 1000 |
[0295] From the above results, it was found that the compound group of the present invention
also has excellent cell growth inhibitory activity against the exon 20 insertion mutant
EGFR expressing cell lines (H1975-EGFRinsSVD and LXF 2478).
Test Example 4 Evaluation of oral absorbability
[0296] The compound of the present invention was suspended or dissolved in 0.5% HPMC aqueous
solution and 0.1 N hydrochloric acid, and the obtained suspension or solution was
orally administered to BALB/cA mice (CLEA Japan, Inc.) at a dose of 50 mg/kg/day.
At 0.5, 1, 2, 4 and 6 hours after completion of the oral administration, blood was
collected from the facial vein over time, so as to obtain plasma. The concentration
of the compound in the obtained plasma was measured by LC-MS/MS, and the oral absorbability
of the present compound was evaluated.
[0297] The results are shown in the following Table 6.
[Table 6]
Example No. |
AUC 0 - 6 hr (µM·hr) |
Example No. |
AUC 0 - 6 hr (µM·hr) |
1 |
50 |
2 |
15 |
3 |
24 |
4 |
12 |
5 |
20 |
6 |
17 |
7 |
15 |
8 |
15 |
9 |
51 |
10 |
50 |
11 |
31 |
12 |
36 |
13 |
18 |
14 |
27 |
15 |
34 |
16 |
15 |
17 |
21 |
18 |
15 |
19 |
6.1 |
Comp. Ex. 2 |
1.5 |
[0298] From the above results, it was found that the compound of the present invention was
contained in a sufficient concentration in the plasma, so that the present compound
exhibited favorable oral absorbability. In contrast, the compound of Comparative Example
2 had oral absorbability that was more than 4 times more attenuated than the compound
of the present invention.
Test Example 5 Evaluation of brain penetration properties
[0299] The compound of the present invention was suspended or dissolved in 0.5% HPMC aqueous
solution and 0.1 N hydrochloric acid, and the obtained suspension or solution was
orally administered to BALB/cA mice (CLEA Japan, Inc.) at a dose of 50 mg/kg/day.
At 0.5 hours after completion of the oral administration, blood was collected from
the facial vein, and whole brain was then excised, so as to obtain plasma and brain
samples. Water was added to the obtained brain sample in 3 times the volume of the
brain sample, and the resultant was then homogenized using an ultrasonic homogenizer,
so as to obtain a brain homogenate. The concentration of the compound in the obtained
plasma and brain homogenate was measured by LC-MS/MS, and the brain penetration properties
of the present compound were evaluated from the brain/plasma concentration of the
compound.
The results are shown in the following Table 7.
[0300]
[Table 7]
Example No. |
Compound concentration in plasma (µM) |
Compound concentration in brain (µM) |
Kp value (Compound concentration in brain/plasma) |
1 |
9.1 |
1.4 |
0.15 |
2 |
6.6 |
1.8 |
0.27 |
3 |
11 |
1.4 |
0.13 |
4 |
8.3 |
2.8 |
0.34 |
5 |
15 |
2.2 |
0.15 |
6 |
7.5 |
1.3 |
0.17 |
7 |
7.9 |
1.1 |
0.14 |
8 |
9.9 |
3.3 |
0.33 |
9 |
13 |
2.4 |
0.18 |
10 |
13 |
2.4 |
0.18 |
11 |
12 |
2.7 |
0.23 |
12 |
11 |
3.2 |
0.29 |
13 |
13 |
2.8 |
0.22 |
14 |
9.9 |
2.1 |
0.21 |
15 |
8.1 |
1.2 |
0.15 |
16 |
12 |
4.4 |
0.35 |
17 |
17 |
6.5 |
0.39 |
18 |
7.7 |
1.6 |
0.22 |
19 |
4.9 |
0.7 |
0.14 |
Comp. Ex. 2 |
1.6 |
0.008 |
0.005 |
[0301] From the above results, it was found that the compound of the present invention had
a high brain/plasma compound concentration (Kp value) and thus, exhibited favorable
brain penetration properties. On the other hand, the brain concentration of the compound
of Comparative Example 2 was more than 80 times more attenuated than that of the compound
of the present invention.
Test Example 6 Antitumor effect confirmation test (in vivo) on subcutaneous transplantation models, into which H1975-EGFRinsSVD cell line is
transplanted
[0302] H1975-EGFRinsSVD cell line was cultured in RPMI-1640 (containing 4.5 g/L glucose,
10 mM HEPES and 1 mM sodium pyruvate) (FUJIFILM Wako Pure Chemical Corporation) medium
containing 10% inactivated fetal bovine serum (FBS) in a 5% CO
2 incubator at 37°C.
[0303] The H1975-EGFRinsSVD cells were resuspended at a concentration of 8 × 10
7 cells/mL in PBS. The cell suspension was subcutaneously transplanted at 8 × 10
6 cells/0.1 mL to the right chest of each 6 week old nude mouse (BALB/cAJcl-nu/nu,
CLEA Japan, Inc.) using a 1 mL syringe for tuberculin and a 25 G injection needle.
[0304] When the tumor volumes of nude mice having tumor engraftment became on the order
of 100 to 200 mm
3, the mice were assigned to groups each involving 6 animals by random stratification
such that the average tumor volume was equal among the groups.
[0305] The test compound used was the compounds of Examples 2, 11, and 12, and a 0.5% HPMC
aqueous solution was used as a control. The compounds of Examples 2, 11 and 12 were
orally administered at doses of 25 mg/kg/day, 25 mg/kg/day and 50 mg/kg/day, respectively.
[0306] Each test compound or the control was orally administered every day for 14 days (Days
1 - 14) from the day following the grouping.
[0307] To compare the transition of tumor growth over time by the administration of each
test compound, a tumor volume (which is also referred to as "TV" below) was measured
at a frequency of twice a week over time. For the measurement of the body weight,
an animal electronic balance was used. A body weight change percentage is also referred
to as "BWC" below. A body weight change percentage on the n
th day (BWCn) from the body weight on the n
th day (BWn) was calculated according to the equation given below. The transition of
average TV and BWC values of the individuals is shown in Figures 1 and 2.
[0308] When the average TV value of the compound administration group on the final evaluation
day (Day 15) was smaller than that of the control group and exhibited statistically
significant difference (Dunnett type multiple comparison test), this compound was
judged as being effective (P < 0.001) and is indicated by the mark * in the drawing.
The results are shown in Figure 1.
[0309] As a result of conducting analysis by the Dunnett type multiple comparison test,
it was shown that the tumor volume was statistically significantly low (P < 0.001)
for all the invention compounds as compared with the control group. From the results
of this test, it was found that the compounds of Examples 2, 11, and 12 have excellent
antitumor effects against the exon 20 insertion mutant EGFR expressing cell line (H1975-EGFRinsSVD)
transplanted into the nude mouse chest. Moreover, a body weight reduction of 20% or
more was not observed in all of the mice to which each compound had been administered.
Test Example 7 Antitumor effect confirmation test (in vivo) on direct brain transplantation models, into which luciferase gene-introduced exon
20 insertion mutant EGFR expressing cell line (H1975-EGFRinsSVD-Luc) is directly transplanted
[0310] The antitumor effects and life-extending effects of the invention compound on direct
brain transplantation models were evaluated using H1975-EGFRinsSVD-Luc line obtained
by introducing luciferase into the human mutant EGFR-introduced cell line H1975-EGFRinsSVD.
[0311] The H1975-EGFRinsSVD-Luc cells used were prepared by introducing pJTI(R) FAST DEST
vector encoding luciferase, together with pJTI(R) PhiC31 Integrase expression vector,
to NCI-H1975-EGFRinsSVD cells by electroporation with Amaxa(R) Cell Line Nucleofector(R)
Kit R, followed by selection using hygromycin B (Nacalai Tesque, Inc.).
[0312] The H1975-EGFRinsSVD-Luc cell line was cultured in RPMI-1640 (containing 4.5 g/L
glucose, 10 mM HEPES and 1 mM sodium pyruvate) (FUJIFILM Wako Pure Chemical Corporation)
medium containing 10% inactivated fetal bovine serum (FBS) in a 5% CO
2 incubator at 37°C.
[0313] The H1975-EGFRinsSVD-Luc cells were resuspended at a concentration of 12.5 × 10
7 cells/mL in PBS.
[0314] Using a mouse ear bar, a nude mouse with 6 to 7 weeks old (BALB/cAJcl-nu/nu, CLEA
Japan, Inc.) was fixed in a brain stereotaxic apparatus, and the skin on the top of
the head was disinfected by the application of an Isodine-containing antiseptic solution
using a sterile cotton swab and was then excised with a surgical knife.
[0315] A microdrill was used to drill a hole in the skull, and then, using a needle, a manipulator,
and a syringe pump, 2 µL of the cell suspension was transplanted into the brain at
a rate of 0.8 µL/min.
[0316] As a reference of the amount of brain tumor, 26 days after the transplantation, Total
Flux (Photon/sec) was measured in all of the survival cases, using IVIS (PerkinElmer,
Inc., model: Lumina II). Based on the obtained results, 10 animals were assigned to
each group by random stratification such that the average total flux was equal among
the groups.
[0317] The test compound used was the compound of Example 11, and a 0.5% HPMC aqueous solution
was used as a control. The compound of Example 11 was administered at a dose of 12.5
mg/kg/day or 25 mg/kg/day.
[0318] The compound of the present invention or the control was orally administered once
a day, every day, for 38 days (Days 27 - 64) from the following day of the grouping
day.
[0319] For judgment of the presence or absence of antitumor effects, the value (Log10) obtained
by logarithmic transformation of the total flux on the antitumor effect judgment day
(Day 47) after 3-week drug administration from the following day (Day 27) of the grouping
day was used.
[0320] A graph was prepared with the value obtained by the average total flux of each group
as a vertical axis, and with the number of days (Day) after the transplantation as
a horizontal axis. The transition of the total flux over time in the drug administration
period was observed.
[0321] For judgment of the presence or absence of life-extending effects, the number of
survival days on the final life-extending effect evaluation day from after cell transplantation
(Days 0 - 65) in the test compound group compared with the control group was analyzed
by the Log-Rank test.
[0322] The results are shown in Figures 3 and 4 given below. The value obtained by logarithmic
transformation (Log10) of the total flux on Day 47 in each group was analyzed by the
Dunnett type multiple comparison test. As a result, it was demonstrated that the aforementioned
value of the test compound group was statistically significantly lower than the value
of the control group (significance level (both sides): 5%) (P < 0.001). From the results
of this test, it was found that the compound of the present invention has antitumor
effects against the exon 20 insertion mutant EGFR expressing cell line (H1975-EGFRinsSVD-Luc)
transplanted into the nude mouse brain.
[0323] In addition, the number of survival days on the final life-extending effect evaluation
day from after cell transplantation (Days 0 - 65) in the test compound group compared
with the control group was analyzed by the Log-Rank test. As a result, statistically
significant life-extending effects were observed, as compared with the control group
(P < 0.05).
[0324] As described above, the compound of the present invention or a salt thereof has
EGFR inhibitory activity and has brain penetration properties and as such, is useful
as an EGFR inhibitor or a therapeutic agent for EGFR-positive tumor.